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Sample records for k45 kd45 kb5

  1. Photoconductivity and dielectric studies of potassium pentaborate crystal (KB5)

    Indian Academy of Sciences (India)

    V Joseph; S Gunasekaran; V Santhanam

    2003-06-01

    Single crystal of potassium pentaborate (KB5) has been grown by solution growth technique. FTIR and laser Raman measurements are carried out to make a qualitative analysis on KB5 crystal. Dielectric behaviour of the KB5 crystal has been studied in the microwave region using K-band microwave bench equipped with the Gunn Oscillator guided with rectangular wave-guide. To confirm the suitability of this crystal as electro optic device, its dielectric behaviour with the change of frequency has also been investigated. Photoconductivity studies were also carried out on this material. It was interesting to observe that the KB5 crystal exhibited negative photoconductivity.

  2. Growth, Optical, Mechanical, Dielectric and Theoretical Studies on Potassium Pentaborate Tetrahydrate (KB_5O_8·4H_2O) Single Crystal by Modified Sankaranarayanan-Ramasamy Method

    Institute of Scientific and Technical Information of China (English)

    C. Justin Raj; S. Krishnan; S. Dinakaran; J. Mary Linet; R. Uthrakumar; R. Robert; Jerome Das

    2009-01-01

    A nonlinear optical single crystal of potassium pentaborate tetrahydrate (KB_5O_8·4H_2O) has been grown from aqueous solution by using unidirectional crystal growth method of Sankaranarayanan-Ramasamy (SR) with a due modification in the growth assembly. Potassium pentaborate crystal of 60 mm length and 10 mm diameter has been grown along (100) plane with a growth rate of 3 mm per day within a period of 20 days. The grown crystal was subjected to single crystal X-ray diffraction analysis to confirm that the crystal belongs to the orthorhombic system. Some fundamental data such as valance electron plasma energy, Penn gap, Fermi energy and electronic polarizability of the grown crystal were calculated. The presence of borate in the grown crystal was confirmed by Fourier transform infrared (FTIR) spectroscopy. The optical transmission property of the grown crystal was analyzed using ultra violet (UV) visible spectral analysis. Surface morphology of the growth plane was observed using scanning electron microscopy (SEM). The mechanical strength of the crystals was found out using Vickers microhardness test along the growth axis. Frequency dependent dielectric constant of the grown crystal was studied for various temperatures along (100) plane.

  3. Structures of three different neutral polysaccharides of Acinetobacter baumannii, NIPH190, NIPH201, and NIPH615, assigned to K30, K45, and K48 capsule types, respectively, based on capsule biosynthesis gene clusters.

    Science.gov (United States)

    Shashkov, Alexander S; Kenyon, Johanna J; Arbatsky, Nikolay P; Shneider, Mikhail M; Popova, Anastasiya V; Miroshnikov, Konstantin A; Volozhantsev, Nikolay V; Knirel, Yuriy A

    2015-11-19

    Neutral capsular polysaccharides (CPSs) were isolated from Acinetobacter baumannii NIPH190, NIPH201, and NIPH615. The CPSs were found to contain common monosaccharides only and to be branched with a side-chain 1→3-linked β-d-glucopyranose residue. Structures of the oligosaccharide repeat units (K units) of the CPSs were elucidated by 1D and 2D (1)H and (13)C NMR spectroscopy. Novel CPS biosynthesis gene clusters, designated KL30, KL45, and KL48, were found at the K locus in the genome sequences of NIPH190, NIPH201, and NIPH615, respectively. The genetic content of each gene cluster correlated with the structure of the CPS unit established, and therefore, the capsular types of the strains studied were designated as K30, K45, and K48, respectively. The initiating sugar of each K unit was predicted, and glycosyltransferases encoded by each gene cluster were assigned to the formation of the linkages between sugars in the corresponding K unit.

  4. Interstitial incorporation of plutonium into a low-dimensional potassium borate.

    Science.gov (United States)

    Wang, Shuao; Diwu, Juan; Simonetti, Antonio; Booth, Corwin H; Albrecht-Schmitt, Thomas E

    2011-11-01

    The molten boric acid flux reaction of PuBr(3) with KBO(2) at 200 °C results in the formation of large light-yellow crystals of K[B(5)O(7)(OH)(2)]·H(2)O:Pu(4+). Single-crystal X-ray diffraction experiments on the Pu-doped K[B(5)O(7)(OH)(2)]·H(2)O demonstrate two features: (1) K[B(5)O(7)(OH)(2)]·H(2)O:Pu(4+) adopts a one-dimensional borate chain structure with void spaces between the chains. (2) The doping plutonium atoms do not reside on the potassium sites. The latter are not fully occupied. Both laser-ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) and energy-dispersive spectrometry analyses indicate that plutonium atoms are uniformly distributed in crystals of K[B(5)O(7)(OH)(2)]·H(2)O with an atomic K:Pu ratio of approximately 65:1 measured by LA-ICP-MS. UV-vis-NIR spectra taken from both freshly made and one day old crystals show that the plutonium present within the crystals is predominantly characterized by Pu(IV). A small amount of Pu(III) is also present initially, but slowly oxidized to Pu(IV) via interaction with oxygen in the air. X-ray absorption near-edge structure and extended X-ray absorption fine structure spectroscopic measurements confirm that plutonium is mainly present as a form similar to that of a PuO(2) cluster. The combined results suggest that the clusters containing Pu(IV) ions are uniformly distributed in the void spaces between the borate chains.

  5. Support for the Forty-Second Annual Gaseous Electronics Conference Held in Palo Alto, California on 16-20 October 1989

    Science.gov (United States)

    1990-05-07

    Lacour, V. Puech, S. Mizzi , S. Pasquiers, and M. Legentil 8:26 - 8:39 FB-3 DETERMINATION OF THE ION TEMPERATURE IN A HeSe-LASER DISCHARGE FROM GAIN...and V. PUECH, S. MIZZI , S. PASQUIERS, M. LEGENTIL, LPGP Universit6 Paris-Sud, FRANCE - The electric properties of X-ray phototriggered discharges in...CA-1 Mitchell, J.B.A. FA-3 Mitchner, M. E-28L Mizzi , S. FB-2 Moeny, W. M. KB-4, KB-5Lacour, B. FB-2 Moore, J. H. BA-3, LB-2 Lagally, M. G. NA-7

  6. STS-37 Pilot Cameron uses SAREX to communicate amateur radio operators

    Science.gov (United States)

    1991-01-01

    STS-37 Pilot Kenneth D. Cameron, wearing Shuttle Amateur Radio Experiment (SAREX) headset (HDST), communicates with amateur radio operators and students while on aft flight deck aboard Atlantis, Orbiter Vehicle (OV) 104. SAREX provided radio transmissions between ground based amateur radio operators around the world and Cameron (call sign KB5AWP) and the other crewmembers, all of whom are licensed amateur radio operators. SAREX enabled students from all over the United States to have a chance to communicate with an astronaut in space. The cloud-covered surface of the Earth is visible above Cameron framed in the overhead window W8.

  7. Morphological, physiological and plant infectivity characterization of Frankia strains isolated from Casuarina’s nodules

    OpenAIRE

    Patrick Moritz; Kelly Campos Guerra P. de Goes; Souza,José Roberto P de; Letícia Trindade Ataíde; Diva Souza Andrade

    2007-01-01

    Frankia are soil microorganisms that form symbiosis with roots of tree species called actinorhizal plants and are capable of fixing atmospheric N2. This study was carried out to characterize morphologically, physiologically and to assess the nodulation of four Frankia reference strains (HFPCcI3, JCT287, KB5 and F59) and 12 (IPRF) isolated from root nodules of Casuarina plants. All strains (Reference and IPRF) were characterized as Gram-positive and 50% as acid-fast. The Frankia strains produc...

  8. Potassium pentaborate

    Directory of Open Access Journals (Sweden)

    Qi Wu

    2011-11-01

    Full Text Available The title compound, K[B5O7(OH2], was obtained from a hydrothermal reaction. The structure is composed of one K+ cation and a polyborate 1∞[B5O7(OH2]− anion, which consists of two six-membered rings linked by a common BO4 tetrahedron. The [B5O7(OH2]− units are linked together through two exocyclic O atoms to neighbouring units, forming a helical chain structure extending parallel to [010]. Adjacent chains are further connected into a three-dimensional structure by K—O bonds and weak O—H...O hydrogen-bond interactions.

  9. Targeted expression of human FSH receptor Asp567Gly mutant mRNA in testis of transgenic mice: role of human FSH receptor promoter

    Institute of Scientific and Technical Information of China (English)

    VerenaNordhoff; JorgGromoll; LucaFoppiani; C.MarcLuetjens; StefanSchlatt; ElenaKostova; IlpoHuhtaniemi; EberhardNieschlag; ManuelaSimoni

    2003-01-01

    Aim:To specifically express the Asp567Gly human follicle-stimulating hormone receptor (FSHR) under the control of its promoter to evaluate the phenotypic consequences in the presence of normal pituitary function.Methods:We produced transgenic mice overexpressing the Asp567Gly human FSHR under the control of a 1.5kb 5’-flanking region fragment of its promoter.Results: Mice were phenotypically normal and fertile.In males,mRNA could be detected in the testis and the brain, indicating that the 1.5kb promoter fragment drives expression not only in the gonads. The testis weight/body weight ratio and the testosterone levels in transgenic and non-transgenic littermates were similar. By in situ hybridisation we found that the transgenic FSHR was highly expressed in Sertoli cells,spermatocytes and round spermatids. However, a radioligand receptor assay failed to show a significant difference in total FSHR binding sites in testis homogenates of transgenic and wild type animals, suggesting that the transgenic FSHR is probably not translated into functional receptor protein. Conclusion: A 1.5kb 5"-region of the human FSHR drives mRNA expression of the transgene in the testis but leads to ectopic expression in germ cells and in the brain. No phenotypic consequences could be documented due to the lack of protein expression.

  10. Isolation and functional characterization of the human 90K promoter

    DEFF Research Database (Denmark)

    Brakebusch, C; Sures, I; Jallal, B;

    1999-01-01

    90K is a secreted protein thought to be involved in the body's defense against pathogens and cancer. To elucidate its transcriptional regulation, the promoter of human 90K (HGMW-approved symbol LGAL S3BP) was isolated and characterized. Analysis of the 3. 3-kb 5'-flanking region revealed that it ......90K is a secreted protein thought to be involved in the body's defense against pathogens and cancer. To elucidate its transcriptional regulation, the promoter of human 90K (HGMW-approved symbol LGAL S3BP) was isolated and characterized. Analysis of the 3. 3-kb 5'-flanking region revealed...... that it is a TATA-less promoter, but neither GC-rich nor dependent on SP1 sites. RNase protection assays detected one major transcription start site (+1) and several minor transcription start sites upstream and downstream. Deletion studies defined a minimal promoter (-103 --> -49) and indirectly suggested positive...... synergism between different elements within it. Consistent with the proposed function of 90K, its promoter activity could be stimulated by poly(I). poly(C), mimicking viral infection. Two regions mediating induction by poly(I). poly(C) (-171 --> -112, -32 --> 46) were identified by deletion mutants. A small...

  11. Domain Modeling: NP_003332.1 [SAHG[Archive

    Lifescience Database Archive (English)

    Full Text Available NP_003332.1 chr3 Crystal structure of human ubiquitin-conjugating enzyme E2 E1 p3bzha_ chr3/NP_003332....1/NP_003332.1_apo_21-193.pdb blast 23E,24K,25E,26T,27N,29P,33E,38M,39S,40K,41N,42S,43K,45

  12. Cutting Edge: RIP1 kinase activity is dispensable for normal development but is a key regulator of inflammation in SHARPIN-deficient mice.

    Science.gov (United States)

    Berger, Scott B; Kasparcova, Viera; Hoffman, Sandy; Swift, Barb; Dare, Lauren; Schaeffer, Michelle; Capriotti, Carol; Cook, Michael; Finger, Joshua; Hughes-Earle, Angela; Harris, Philip A; Kaiser, William J; Mocarski, Edward S; Bertin, John; Gough, Peter J

    2014-06-15

    RIP1 (RIPK1) kinase is a key regulator of TNF-induced NF-κB activation, apoptosis, and necroptosis through its kinase and scaffolding activities. Dissecting the balance of RIP1 kinase activity and scaffolding function in vivo during development and TNF-dependent inflammation has been hampered by the perinatal lethality of RIP1-deficient mice. In this study, we generated RIP1 kinase-dead (Ripk1(K45A)) mice and showed they are viable and healthy, indicating that the kinase activity of RIP1, but not its scaffolding function, is dispensable for viability and homeostasis. After validating that the Ripk1(K45A) mice were specifically protected against necroptotic stimuli in vitro and in vivo, we crossed them with SHARPIN-deficient cpdm mice, which develop severe skin and multiorgan inflammation that has been hypothesized to be mediated by TNF-dependent apoptosis and/or necroptosis. Remarkably, crossing Ripk1(K45A) mice with the cpdm strain protected against all cpdm-related pathology. Together, these data suggest that RIP1 kinase represents an attractive therapeutic target for TNF-driven inflammatory diseases.

  13. Epistemic and doxastic logic with restrictions Lógicas epistémica y doxástica con restricciones

    Directory of Open Access Journals (Sweden)

    Manuel Sierra A

    2010-12-01

    Full Text Available Are presented as extensions of classical propositional calculus hierarchies of deductive systems LDR–n and LER–n with n > 1. LER–n is the epistemic logic with restrictions, LDR–n is the doxastic logic with restrictions. The systems LER–1 and LDR–1 are the classical propositional calculus. System LER–(n + 1 can be seen as the result of applying the rule: if X is theorem of LER–n then +X is theorem of LER–(n + 1. Systems also restricts the validity of the axioms +(X → Y → (+X → +Y and +X → X, in terms of depth (complexity with respect to the operator + of X and Y , and also includes restricted versions of the axioms of positive and negative introspection. LER system results from the union of LER–n systems, and can be seen as the S5 modal logic system with different types of restrictions. Changing +X → X by +X →∼+∼X are built LDR–n and the LDR systems. LDR can be seen as the KD45 modal logic system with different types of restrictions. The systems are characterized with a embedded worlds semantics, with which the ‘omniscience logical problem’ is limited.Se presentan como extensiones del cálculo proposicional clásico las jerarquías de sistemas deductivos LER–n y LDR–n, con n ≥ 1. LER–n es la Lógica epistémica con restricciones de profundidad–n, LDR–n es la Lógica doxástica con restricciones de profundidad–n. Los sistemas LER–1 y LDR–1 son el cálculo proposicional clásico. El sistema LER–(n + 1 puede ser visto como el resultado de aplicar la regla: de X se infiere +X, una vez a los teoremas del sistema LER–n, además, se restringe la validez de los axiomas +(X → Y → (+X → +Y y +X → X en términos de la profundidad (complejidad respecto al operador + de X y de Y , y también se incluyen versiones generalizadas y con restricciones de los axiomas de introspección positiva y negativa. El sistema LER resulta de la reunión de los sistemas de la jerarquía, y puede ser visto

  14. Discovery of Aptamer Ligands for Hepatic Stellate Cells Using SELEX.

    Science.gov (United States)

    Chen, Zhijin; Liu, Hao; Jain, Akshay; Zhang, Li; Liu, Chang; Cheng, Kun

    2017-01-01

    Insulin like growth factor II receptor (IGFIIR) is a transmembrane protein overexpressed in activated hepatic stellate cells (HSCs), which are the major target for the treatment of liver fibrosis. In this study, we aim to discover an IGFIIR-specific aptamer that can be potentially used as a targeting ligand for the treatment and diagnosis of liver fibrosis. Systematic evolution of ligands by exponential enrichment (SELEX) was conducted on recombinant human IGFIIR to identify IGFIIR-specific aptamers. The binding affinity and specificity of the discovered aptamers to IGFIIR and hepatic stellate cells were studied using flow cytometry and Surface Plasmon Resonance (SPR). Aptamer-20 showed the highest affinity to recombinant human IGFIIR protein with a Kd of 35.5 nM, as determined by SPR. Aptamer-20 also has a high affinity (apparent Kd 45.12 nM) to LX-2 human hepatic stellate cells. Binding of aptamer-20 to hepatic stellate cells could be inhibited by knockdown of IGFIIR using siRNA, indicating a high specificity of the aptamer. The aptamer formed a chimera with an anti-fibrotic PCBP2 siRNA and delivered the siRNA to HSC-T6 cells to trigger silencing activity. In Vivo biodistribution study of the siRNA-aptamer chimera also demonstrated a high and specific uptake in the liver of the rats with CCl4-induced liver fibrosis. These data suggest that aptamer-20 is a high-affinity ligand for antifibrotic and diagnostic agents for liver fibrosis.

  15. HindIII identifies a two allele DNA polymorphism of the human cannabinoid receptor gene (CNR)

    Energy Technology Data Exchange (ETDEWEB)

    Caenazzo, L.; Hoehe, M.R.; Hsieh, W.T.; Berrettini, W.H.; Bonner, T.I.; Gershon, E.S. (National Inst. of Health, Bethesda, MD (United States))

    1991-09-11

    HCNR p5, a 0.9 kb BamHI/EcoRI fragment from the human cannabinoid receptor gene inserted into pUC19, was used as probe. The fragment is located in an intron approximately 14 kb 5{prime} of the initiation codon. This fragment is a clean single copy sequence by genomic blotting. Hybridization of human genomic DNA digested with HindIII identified a two allele RFLP with bands at 5.5 (A1) and 3.3 kb (A2). The human cannabinoid receptor gene has been genetically mapped in CEPH reference pedigrees to the centromeric/q region of chromosome 6. In situ hybridization localizes it to 6q14-q15. Codominant segregation has been observed in 26 informative two- and three-generation CEPH pedigrees and in 14 medium-sized disease families.

  16. Synthesis of dihydromyricetin-manganese (II) complex and interaction with DNA

    Science.gov (United States)

    Guo, Qingquan; Yuan, Juan; Zeng, Jinhua; He, Xiangzhu; Li, Daguang

    2012-11-01

    Dihydromyricetin has many physiological functions and its metal complex could have better effects. DNA is very important in biological body, but little attention has been devoted to the relationship between dihydromyricetin-metal complex and the DNA. In this paper, dihydromyricetin-Mn (II) complex has been prepared and characterized using UV-vis absorption spectrophotometry, IR spectroscopy, elemental analysis, and thermal gravimetric analysis (TG-DTA Analysis). The interaction of dihydromyricetin-Mn (II) complex with DNA was investigated using UV-vis spectra, fluorescence measurements and viscosity measurements. The results indicate that the dihydromyricetin-manganese (II) complex can intercalate into the stacked base pairs of DNA with binding constant Kb = 5.64 × 104 M and compete with the strong intercalator ethidium bromide for the intercalative binding sites with Stern-Volmer quenching constant, Ksq = 1.16.

  17. Dicty_cDB: VFK294 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available for mK... 45 0.003 ( P59438 ) RecName: Full=Hermansky-Pudlak syndrome 5 protein homol... 45 0.003 AF534398_...la melanogaster RE38137 fu... 43 0.021 (Q297N8) RecName: Full=Hermansky-Pudlak syndrome 5 protein homol... 4...0.082 AB463066_1( AB463066 |pid:none) Synthetic construct DNA, clone: pF... 41 0.082 (Q9UPZ3) RecName: Full=Herman

  18. The Binding Characterization of Cry Insecticidal Proteins to the Brush Border Membrane Vesicles of Helicoverpa armigera, Spodoptera exigua, Spodoptera litura and Agrotis ipsilon

    Institute of Scientific and Technical Information of China (English)

    LU Qiong; CAO Guang-chun; ZHANG Li-li; LIANG Ge-mei; GAO Xi-wu; ZHANG Yong-jun; GUO Yu-yuan

    2013-01-01

    Cry toxins produced by Bacillus thuringiensis (Bt) are effective biological insecticides against certain insect species. However, there are potential risks of the evolved resistance of insects to Cry toxin owing to decreased binding of toxins to target sites in the brush border membranes of the larva midgut. The Cry toxins with different binding sites in the larval midgut have been considered to be a good combination to deploy in delaying resistance evolution. Bioassay results demonstrated that the toxicity of different Cry toxins ranked differently for each species. The toxicity ranking was Cry1Ac>Cry1Ab>Cry2Ab for Helicoverpa armigera, Cry1B>Cry1C>Cry2Ab for Spodoptera exigua, and Cry2Ab>Cry1B>Cry1C for S. litura. Only Cry2Ab was toxic to Agrotis ipsilon. Binding experiments were performed with 125I-Cry1Ab, 125I-Cry1Ac, 125I-Cry1B, 125I-Cry1C, 125I-Cry2Ab and the brush border membranes vesicles (BBMV) from H. armigera, S. exigua, S. litura and A. ipsilon. The binding of Cry1Ab and Cry1Ac was shown to be saturable by incubating with increasing concentrations of H. armigera BBMV (Kd=(45.00±2.01) nmol L-1 and (12.80±0.18) nmol L-1, respectively;Bmax=(54.95±1.79) ng and (55.44±0.91) ng, separately). The binding of Cry1B was shown to be saturable by incubating with increasing concentrations of S. exigua BBMV (Kd=(23.26±1.66) nmol L-1;Bmax=(65.37±1.87) ng). The binding of 125I-Cry toxins was shown to be non-saturable by incubating with increasing concentrations of S. litura and A. ipsilon BBMV. In contrast, Cry1B and Cry1C showed some combination with the BBMV of S. litura, and a certain amount of Cry2Ab could bind to the BBMV of A. ipsilon. These observations suggest that a future strategy could be devised for the focused combination of specific cry genes in transgenic crops to control target pests, widen the spectrum of insecticide effectiveness and postpone insect resistance evolution.

  19. Kinetics of the initial steps in the aqueous oxidation of thiosulfate by chlorine dioxide.

    Science.gov (United States)

    Pan, Changwei; Stanbury, David M

    2014-08-28

    The reaction of ClO2(•) with S2O3(2-) in aqueous solution is a component of the "crazy clock" reaction of ClO2(-) with S2O3(2-), and under conditions of excess S2O3(2-) the absorbance at 360 nm due to ClO2(•) decays with sigmoidal kinetics. A chain reaction mechanism is inferred on the basis that very small concentrations of SO3(2-) accelerate the reaction, and methionine inhibits the reaction. Pseudo-first-order kinetics is observed in the presence of relatively large methionine concentrations, leading to the simple rate law -d[ClO2]/dt = (ka[S2O3(2-)] + kb[S2O3(2-)](2))[ClO2], with ka = 452 ± 16 M(-1) s(-1) and kb = (5.7 ± 0.2) × 10(5) M(-2) s(-1) at 25 °C and pH 7.6. Under these conditions, the initial products are ClO2(-) and S4O6(2-). A classical electron-transfer mechanism is assigned to the reaction that occurs under conditions of methionine inhibition.

  20. Transcriptional analysis of the DNA polymerase gene of shrimp white spot syndrome virus.

    Science.gov (United States)

    Chen, Li-Li; Wang, Han-Ching; Huang, Chiu-Jung; Peng, Shao-En; Chen, Yen-Gu; Lin, Shin-Jen; Chen, Wei-Yu; Dai, Chang-Feng; Yu, Hon-Tsen; Wang, Chung-Hsiung; Lo, Chu-Fang; Kou, Guang-Hsiung

    2002-09-15

    The white spot syndrome virus DNA polymerase (DNA pol) gene (WSSV dnapol) has already been tentatively identified based on the presence of highly conserved motifs, but it shows low overall homology with other DNA pols and is also much larger (2351 amino acid residues vs 913-1244 aa). In the present study we perform a transcriptional analysis of the WSSV dnapol gene using the total RNA isolated from WSSV-infected shrimp at different times after infection. Northern blot analysis with a WSSV dnapol-specific riboprobe found a major transcript of 7.5 kb. 5'-RACE revealed that the major transcription start point is located 27 nucleotides downstream of the TATA box, at the nucleotide residue A within a CAGT motif, one of the initiator (Inr) motifs of arthropods. In a temporal expression analysis using differential RT-PCR, WSSV dnapol transcripts were detected at low levels at 2-4 h.p.i., increased at 6 h.p.i., and remained fairly constant thereafter. This is similar to the previously reported transcription patterns for genes encoding the key enzyme of nucleotide metabolism, ribonucleotide reductase. Phylogenetic analysis showed that the DNA pols from three different WSSV isolates form an extremely tight cluster. In addition, similar to an earlier phylogenetic analysis of WSSV protein kinase, the phylogenetic tree of viral DNA pols further supports the suggestion that WSSV is a distinct virus (likely at the family level) that does not belong to any of the virus families that are currently recognized.

  1. DNA repair gene polymorphisms in relation to chromosome aberration frequencies in retired radiation workers

    Energy Technology Data Exchange (ETDEWEB)

    Wilding, Craig S. [Genetics Department, Westlakes Research Institute, Westlakes Science and Technology Park, Moor Row, Cumbria CA24 3JY (United Kingdom)]. E-mail: craig.wilding@westlakes.ac.uk; Relton, Caroline L. [Genetics Department, Westlakes Research Institute, Westlakes Science and Technology Park, Moor Row, Cumbria CA24 3JY (United Kingdom); Paediatric and Lifecourse Epidemiology Research Group, School of Clinical Medical Sciences (Child Health), Newcastle University, Sir James Spence Institute, Royal Victoria Infirmary, Newcastle-upon-Tyne NE1 4LP (United Kingdom); Rees, Gwen S. [Genetics Department, Westlakes Research Institute, Westlakes Science and Technology Park, Moor Row, Cumbria CA24 3JY (United Kingdom); Tarone, Robert E. [International Epidemiology Institute, 1455 Research Boulevard, Suite 550, Rockville, MD 20850 (United States); Whitehouse, Caroline A. [Genetics Department, Westlakes Research Institute, Westlakes Science and Technology Park, Moor Row, Cumbria CA24 3JY (United Kingdom); Tawn, E. Janet [Genetics Department, Westlakes Research Institute, Westlakes Science and Technology Park, Moor Row, Cumbria CA24 3JY (United Kingdom)

    2005-02-15

    Polymorphic variation in DNA repair genes was examined in a group of retired workers from the British Nuclear Fuels plc facility at Sellafield in relation to previously determined translocation frequencies in peripheral blood lymphocytes. Variation at seven polymorphisms in four genes involved in the base excision repair (XRCC1 R194W, R399Q and a [AC]{sub n} microsatellite in the 3' UTR) and double strand break repair (XRCC3 T241M and a [AC]{sub n} microsatellite in intron 3 of XRCC3, XRCC4 I134T, and a GACTAn microsatellite located 120kb 5' of XRCC5) pathways was determined for 291 retired radiation workers who had received cumulative occupational external radiation doses of between 0 and 1873mSv. When the interaction between radiation dose and each DNA repair gene polymorphism was examined in relation to translocation frequency there was no evidence for any of the polymorphisms studied influencing the response to occupational exposure. A positive interaction observed between genotype (individuals with at least one allele >=20 repeat units) at a microsatellite locus in the XRCC3 gene and smoking status should be interpreted cautiously because interactions were investigated for seven polymorphisms and two exposures. Nonetheless, further research is warranted to examine whether this DNA repair gene variant might be associated with a sub-optimal repair response to smoking-induced DNA damage and hence an increased frequency of translocations.

  2. Distal Interleukin-1β (IL-1β) Response Element of Human Matrix Metalloproteinase-13 (MMP-13) Binds Activator Protein 1 (AP-1) Transcription Factors and Regulates Gene Expression*

    Science.gov (United States)

    Schmucker, Adam C.; Wright, Jason B.; Cole, Michael D.; Brinckerhoff, Constance E.

    2012-01-01

    The collagenase matrix metalloproteinase-13 (MMP-13) plays an important role in the destruction of cartilage in arthritic joints. MMP-13 expression is strongly up-regulated in arthritis, largely because of stimulation by inflammatory cytokines such as IL-1β. Treatment of chondrocytes with IL-1β induces transcription of MMP-13 in vitro. IL-1β signaling converges upon the activator protein-1 transcription factors, which have been shown to be required for IL-1β-induced MMP-13 gene expression. Using chromatin immunoprecipitation (ChIP), we detected activator protein-1 binding within an evolutionarily conserved DNA sequence ∼20 kb 5′ relative to the MMP-13 transcription start site (TSS). Also using ChIP, we detected histone modifications and binding of RNA polymerase II within this conserved region, all of which are consistent with transcriptional activation. Chromosome conformation capture indicates that chromosome looping brings this region in close proximity with the MMP-13 TSS. Finally, a luciferase reporter construct driven by a component of the conserved region demonstrated an expression pattern similar to that of endogenous MMP-13. These data suggest that a conserved region at 20 kb upstream from the MMP-13 TSS includes a distal transcriptional response element of MMP-13, which contributes to MMP-13 gene expression. PMID:22102411

  3. Transcriptional Regulation of Expression of the Maize Aldehyde Dehydrogenase 7 Gene (ZmALDH7B6) in Response to Abiotic Stresses

    Institute of Scientific and Technical Information of China (English)

    GU Ri-liang

    2014-01-01

    Aldehyde dehydrogenases (ALDHs) represent a large protein family, which includes several members that catalyze the oxidation of an aldehyde to its corresponding carboxylic acid in plants. Genes encoding members of theALDH7 subfamily have been suggested to play important roles in various stress adaptations in plants. In this study, quantitative RT-PCR analysis revealed that a maizeALDH7 subfamily member (ZmALDH7B6) was constitutively expressed in various organs, including roots, leaves, immature ears, tassels, and developing seeds. The abundance ofZmALDH7B6 mRNA transcripts in maize roots was increased by ammonium, NaCl, and mannitol treatments. To further analyze tissue-speciifc and stress-induced expression patterns, the 1.5-kb 5´-lfankingZmALDH7B6 promoter region was fused to the β-glucuronidase (GUS) reporter gene and introduced into maize plants. In roots of independent transgenic lines, there was signiifcant induction of GUS activity in response to ammonium supply, conifrming ammonium-dependent expression ofZmALDH7B6 at the transcript level. Histochemical staining showed that GUS activity driven by theZmALDH7B6 promoter was mainly localized in the vascular tissues of maize roots. These results suggested thatZmALDH7B6 is induced by multiple environmental stresses in maize roots, and may play a role in detoxifying aldehydes, particularly in vascular tissue.

  4. Seminal vesicles and urinary bladder as sites of aromatization of androgens in men, evidenced by a CYP19A1-driven luciferase reporter mouse and human tissue specimens.

    Science.gov (United States)

    Strauss, Leena; Rantakari, Pia; Sjögren, Klara; Salminen, Anu; Lauren, Eve; Kallio, Jenny; Damdimopoulou, Pauliina; Boström, Minna; Boström, Peter J; Pakarinen, Pirjo; Zhang, FuPing; Kujala, Paula; Ohlsson, Claes; Mäkelä, Sari; Poutanen, Matti

    2013-04-01

    The human CYP19A1 gene is expressed in various tissues by the use of tissue-specific promoters, whereas the rodent cyp19a1 gene is expressed mainly in the gonads and brain. We generated a transgenic mouse model containing a >100-kb 5' region of human CYP19A1 gene connected to a luciferase reporter gene. The luciferase activity in mouse tissues mimicked the CYP19A1 gene expression pattern in humans. Interestingly, the reporter gene activity was 16 and 160 times higher in the urinary bladder and seminal vesicles, respectively, as compared with the activity in the testis. Accordingly, CYP19A1 gene and P450arom protein expression was detected in those human tissues. Moreover, the data revealed that the expression of CYP19A1 gene is driven by promoters PII, I.4, and I.3 in the seminal vesicles, and by promoters PII and I.4 in the urinary bladder. Furthermore, the reporter gene expression in the seminal vesicles was androgen dependent: Castration decreased the expression ∼20 times, and testosterone treatment restored it to the level of an intact mouse. This reporter mouse model facilitates studies of tissue-specific regulation of the human CYP19A1 gene, and our data provide evidence for seminal vesicles as important sites for estrogen production in males.

  5. Down-Regulation of the Na+-Coupled Phosphate Transporter NaPi-IIa by AMP-Activated Protein Kinase

    Directory of Open Access Journals (Sweden)

    Miribane Dërmaku-Sopjani

    2013-11-01

    Full Text Available Background/Aims: The Na+-coupled phosphate transporter NaPi-IIa is the main carrier accomplishing renal tubular phosphate reabsorption. It is driven by the electrochemical Na+ gradient across the apical cell membrane, which is maintained by Na+ extrusion across the basolateral cell membrane through the Na+/K+ ATPase. The operation of NaPi-IIa thus requires energy in order to avoid cellular Na+ accumulation and K+ loss with eventual decrease of cell membrane potential, Cl- entry and cell swelling. Upon energy depletion, early inhibition of Na+-coupled transport processes may delay cell swelling and thus foster cell survival. Energy depletion is sensed by the AMP-activated protein kinase (AMPK, a serine/threonine kinase stimulating several cellular mechanisms increasing energy production and limiting energy utilization. The present study explored whether AMPK influences the activity of NAPi-IIa. Methods: cRNA encoding NAPi-IIa was injected into Xenopus oocytes with or without additional expression of wild-type AMPK (AMPKα1-HA+AMPKβ1-Flag+AMPKγ1-HA, of inactive AMPKαK45R (AMPKα1K45R+AMPKβ1-Flag+AMPKγ1-HA or of constitutively active AMPKγR70Q (AMPKα1-HA+AMPKβ1-Flag+AMPKγ1R70Q. NaPi-IIa activity was estimated from phosphate-induced current in dual electrode voltage clamp experiments. Results: In NaPi-IIa-expressing, but not in water-injected Xenopus oocytes, the addition of phosphate (1 mM to the extracellular bath solution generated a current (Ip, which was significantly decreased by coexpression of wild-type AMPK and of AMPKγR70Q but not of AMPKαK45R. The phosphate-induced current in NaPi-IIa- and AMPK-expressing Xenopus ooocytes was significantly increased by AMPK inhibitor Compound C (20 µM. Kinetic analysis revealed that AMPK significantly decreased the maximal transport rate. Conclusion: The AMP-activated protein kinase AMPK is a powerful regulator of NaPi-IIa and thus of renal tubular phosphate transport.

  6. The thermodynamic stability and hydration enthalpy of strontium cerate doped by yttrium

    OpenAIRE

    Matskevich, N. I.; Wolf, Th.; Matskevich, M. Yu.

    2005-01-01

    The standard molar enthalpy of formation of SrY0.05Ce0.95O2.975 has been derived by combining the enthalpy of solution of this compound in 1 M HCl + 0.1 KI obtained by us and auxiliary literature data. The following value has been derived: DfH (SrY0.05Ce0.95O2.975, s, 298.15 K) = -1720.4 (3.4) kJ/mol. The obtained value has been used to obtain the formation enthalpy of SrY0.05Ce0.95O2.975 from the mixture of binary oxides (DoxH (298.15 K) = -45.9 (3.4) kJ/mol) and formation enthalpy of reacti...

  7. Numerical simulations of expanding supershells in dwarf irregular galaxies. II. Formation of giant HI rings

    CERN Document Server

    Vorobyov, E I; Basu, Shantanu

    2004-01-01

    We perform numerical hydrodynamic modeling of various physical processes that can form an HI ring as is observed in Holmberg I. Three energetic mechanisms are considered: multiple supernova explosions (SNe), a hypernova explosion associated with a gamma ray burst (GRB), and the vertical impact of a high velocity cloud (HVC). The total released energy has an upper limit of 10^54 ergs. We find that multiple SNe are in general more effective in producing shells that break out of the disk than a hypernova explosion of the same total energy. As a consequence, multiple SNe form rings with a high ring-to-center contrast K 45 deg) the HI image is characterized by two kidney-shaped density enhancements and a mild central depression.

  8. Dicty_cDB: Contig-U11332-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 265_1( AK129265 |pid:none) Mus musculus premature mRNA for mK... 45 0.015 ( P59438 ) RecName: Full=Hermansky....28 (Q297N8) RecName: Full=Hermansky-Pudlak syndrome 5 protein homol... 41 0.28 A... Synthetic construct DNA, clone: pF... 41 0.37 (Q9UPZ3) RecName: Full=Hermansky-Pudlak syndrome 5 protein; A...40 |pid:none) Homo sapiens Hermansky-Pudlak synd... 41 0.37 AK292436_1( AK292436 |pid:none) Homo sapiens cDN...m 3D7 chromo... 38 3.1 BC082542_1( BC082542 |pid:none) Mus musculus Hermansky-Pudlak synd... 37 4.1 AL844509

  9. Development and Building of Radioactive Concrete Pads for calibration of the airborne and ground gamma-ray spectrometers, used in mineral exploration and hydrocarbons; Desenvolvimento e construcao de blocos de concreto radioativo para calibracao de espectrometros gama portateis e aerotransportados, utilizados em exploracao mineral e de hidrocarbonetos

    Energy Technology Data Exchange (ETDEWEB)

    Carlos, Dionisio Uendro

    2006-07-01

    Eight transportable calibration pads were built in to be used as concentration standards for portable and airborne gamma-ray spectrometers calibrations. The pads construction procedure is described in full detail. The pads, with dimensions of 1 m x 1 m x 0,30 m and masses between 593 kg and 673 kg were made radioactive by the addition of different amounts of k-feldspar, caldasite and monazitic sand to the concrete masses. The potassium, uranium and thorium concentration vary significantly in the pads, reaching maximum values of 5,7% of K, 45,6 ppm eU and 137 ppm eTh. The distribution of the gamma radiation flux from the pads surfaces and the heterogeneity magnitudes of the radioactive elements concentration were experimentally established. An example of gamma-ray spectrometer calibration is presented. (author)

  10. Capsular types of Klebsiella pneumoniae revisited by wzc sequencing.

    Directory of Open Access Journals (Sweden)

    Yi-Jiun Pan

    Full Text Available Capsule is an important virulence factor in bacteria. A total of 78 capsular types have been identified in Klebsiella pneumoniae. However, there are limitations in current typing methods. We report here the development of a new genotyping method based on amplification of the variable regions of the wzc gene. Fragments corresponding to the variable region of wzc were amplified and sequenced from 76 documented capsular types of reference or clinical strains. The remaining two capsular types (reference strains K15 and K50 lacked amplifiable wzc genes and were proven to be acapsular. Strains with the same capsular type exhibited ≧94% DNA sequence identity across the variable region (CD1-VR2-CD2 of wzc. Strains with distinct K types exhibited <80% DNA sequence identity across this region, with the exception of three pairs of strains: K22/K37, K9/K45, and K52/K79. Strains K22 and K37 shared identical capsular polysaccharide synthesis (cps genes except for one gene with a difference at a single base which resulted in frameshift mutation. The wzc sequences of K9 and K45 exhibited high DNA sequence similarity but possessed different genes in their cps clusters. K52 and K79 exhibited 89% wzc DNA sequence identity but were readily distinguished from each other at the DNA level; in contrast, strains with the same capsular type as K52 exhibited 100% wzc sequence identity. A total of 29 strains from patients with bacteremia were typed by the wzc system. wzc DNA sequences confirmed the documented capsular type for twenty-eight of these clinical isolates; the remaining strain likely represents a new capsular type. Thus, the wzc genotyping system is a simple and useful method for capsular typing of K. pneumoniae.

  11. GTP binding to the. beta. -subunit of tubulin is greatly reduced in Alzheimers disease

    Energy Technology Data Exchange (ETDEWEB)

    Khatoon, S.; Slevin, J.T.; Haley, B.E.

    1987-05-01

    A decrease occurs (80-100%) in the (/sup 32/P)8N/sub 3/GTP photoinsertion into a cytosolic protein (55K M/sub r/) of Alzheimer's (AD) brain, tentatively identified as the ..beta..-subunit of tubulin (co-migration with purified tubulin, concentration dependence of interaction with GTP, ATP and their 8-azido photoprobes, and similar effects of Ca/sup 2 +/ and EDTA on photoinsertion). This agrees with prior observations of (/sup 32/P)8N/sub 3/GTP interactions with brain tubulin and a recent report on faulty microtubular assembly in AD brain. The decrease in (/sup 32/P)8N/sub 3/GTP photoinsertion into the 55K M/sub r/ protein of AD brain was in contrast with other photolabeled proteins, which remained at equal levels in AD and age-matched normal brain tissues. The 55K and 45K M/sub r/ were the two major (/sup 32/P)8N/sub 3/GTP photoinsertion species in non-AD brain. Of 5 AD brains, the photoinsertion of (/sup 32/P)8N/sub 3/GTP into the 55K M/sub r/ region was low or absent in 4 (55K/45K=0.1); one was 75% below normals (55K/45K=0.24). Total protein migrating at 55K M/sub r/ was similar in AD and controls. AD brain tubulin, while present, has its exchangeable GTP binding site on ..beta..-tubulin blocked/modified such that (/sup 32/P)8N/sub 3/GTP cannot interact normally with this site.

  12. THAP and ATF-2 regulated sterol carrier protein-2 promoter activities in the larval midgut of the yellow fever mosquito, Aedes aegypti.

    Directory of Open Access Journals (Sweden)

    Rong Peng

    Full Text Available Expression of sterol carrier protein-2 (SCP-2 in Aedes aegypti shows a distinct temporal/spatial pattern throughout the life cycle. In order to identify the transcription factors responsible for the larval temporal/spatial regulation of AeSCP-2 transcription, AeSCP-2 promoter activities were studied in vivo via transient transfection of promoter/reporter gene assays. Regulatory sequences upstream -1.3 kb of the transcription start site of AeSCP-2 were found to be critical for the in vivo temporal/spatial promoter activity. Interestingly, the -1.6 kb promoter sequence efficiently drove the larval midgut-specific siRNA expression, indicating that the -1.6 kb upstream sequence is sufficient for temporal/spatial AeSCP-2 transcriptional activity. Four transcription factors were identified in the midgut nuclear extract from feeding larvae via labeled -1.6/-1.3 kb DNA probe pull-down and proteomic analysis. Co-transfection of the promoter/reporter gene with inducible siRNA expression of each transcription factor was performed to confirm the regulatory function of individual transcription factor on AeSCP-2 transcriptional activities in the larval midgut. The results indicate that two of the identified transcription factors, Thanatos-associated protein (THAP and activating transcription factor-2 (ATF-2, antagonistically control AeSCP-2 transcriptional activity in the midgut of feeding larvae via the regulatory sequences between -1.6 to -1.3 kb 5' upstream of the transcription start site. In vivo expression knockdown of THAP and ATF-2 resulted in significant changes in developmental progression, which may be partially due to their effects on AeSCP-2 expression.

  13. Molecular structural and functional characterization of STAT1 gene regulatory region in teleost Channa argus.

    Science.gov (United States)

    Jia, Weizhang; Zhou, Xiuxia

    2010-05-15

    The transcription factor STAT1 is involved in signal transduction of type I and II interferons (IFNs). However, the molecular characteristics of the STAT1 regulatory region still remain to be elucidated in teleosts. In the present study, the complete cDNA and the regulatory region of the STAT1 gene were isolated from snakehead (Channa argus). More than 2.4kb 5'-flanking region of STAT1 shares the regulatory elements of IFN-stimulated response element (ISRE) and IFN-gamma activation site (GAS). Consensus ISRE and GAS were located from -373 to -361 and -716 to -724 in the promoter region, respectively. Moreover, it is noticeable that the crucial elements of ISRE (+698 to +710) and GAS (+294 and +301) are present in the first intron of snakehead STAT1. Comparisons of six vertebrate STAT1 5'-flanking regions all present the common sequence characteristics of IFN-induced gene promoter, which include ISRE, GAS and Sp1 sites. In order to further characterize the snakehead STAT1 regulatory region, six reporter constructs of snakehead STAT1 promoter and first intron were generated to examine the specificity to human interferon-gamma (hIFN-gamma). Only those constructs containing the ISRE element showed notable reporter activity after stimulation of Hela cells with hIFN-gamma. However, sequential deletions of putative transcription factor binding sites indicated that GAS elements have little effect on the promoter and intronic activity in response to hIFN-gamma. Taken together, these results suggest that the regulatory mechanisms of IFN-signalling appear to be mediated in a similar manner in fish and mammals.

  14. Diabetes susceptibility at IDDM2 cannot be positively mapped to the VNTR locus of the insulin gene.

    Science.gov (United States)

    Doria, A; Lee, J; Warram, J H; Krolewski, A S

    1996-05-01

    An inconsistency has come to light between the conclusion of Lucassen et al. that IDDM2 (11p15.5) must lie within a 4.1 kilobase (kb) segment at the insulin (INS) locus and their own data showing statistically significant associations between insulin-dependent diabetes mellitus (IDDM) and markers beyond the boundaries of that segment. We present data from an independent study of 201 IDDM patients and 107 non-diabetic control subjects that also show significant association with a marker 5' of the INS locus. Patients and control subjects were genotyped at INS/+ 1140 A/C (a surrogate for the variable number tandem repeat (VNTR) polymorphism in the regulatory part of the INS gene) and a marker 5' of the tyrosine hydroxylase (TH) gene, TH/pINS500-RsaI, making it 10 kb 5' of the VNTR. Homozygotes for INS/ + 1140 allele '+' were significantly more frequent among IDDM patients than among control subjects (73 vs 45%, p < 0.001) giving an odds ratio of 3.3 (95% confidence interval (CI): 2.0-5.3). A very similar association was found for homozygotes for the TH/RsaI allele '+' (53 vs 31%, p < 0.001) giving an odds ratio of 2.6 (95%CI 1.6-4.2). By multilocus analysis, the TH/RsaI allele '+' identified a subset of INS/ + 1140 alleles '+' haplotypes that are more specifically associated with IDDM (odds ratio = 5.4, 95%CI 2.9-10.4) than allele + 1140 '+' as a whole. In conclusion, the segment of chromosome 11 that is associated with IDDM spans, at least, the INS and TH loci. No legitimate claim can be made that IDDM2 corresponds to the VNTR polymorphism at the INS locus until the correct boundaries for IDDM2 have been defined and other loci within them have been excluded as determinants of IDDM.

  15. Deep resequencing of CFTR in 762 F508del homozygotes reveals clusters of non-coding variants associated with cystic fibrosis disease traits

    Science.gov (United States)

    Vecchio-Pagán, Briana; Blackman, Scott M; Lee, Melissa; Atalar, Melis; Pellicore, Matthew J; Pace, Rhonda G; Franca, Arianna L; Raraigh, Karen S; Sharma, Neeraj; Knowles, Michael R; Cutting, Garry R

    2016-01-01

    Extensive phenotypic variability is commonly observed in individuals with Mendelian disorders, even among those with identical genotypes in the disease-causing gene. To determine whether variants within and surrounding CFTR contribute to phenotypic variability in cystic fibrosis (CF), we performed deep sequencing of CFTR in 762 patients homozygous for the common CF-causing variant, F508del. In phase 1, ~200 kb encompassing CFTR and extending 10 kb 5′ and 5 kb 3′ of the gene was sequenced in 486 F508del homozygotes selected from the extremes of sweat chloride concentration. In phase 2, a 510 kb region, which included the entire topologically associated domain of CFTR, was sequenced in 276 F508del homozygotes drawn from extremes of lung function. An additional 163 individuals who carried F508del and a different CF-causing variant were sequenced to inform haplotype construction. Region-based burden testing of both common and rare variants revealed seven regions of significance (α=0.01), five of which overlapped known regulatory elements or chromatin interactions. Notably, the −80 kb locus known to interact with the CFTR promoter was associated with variation in both CF traits. Haplotype analysis revealed a single rare recombination event (1.9% frequency) in intron 15 of CFTR bearing the F508del variant. Otherwise, the majority of F508del chromosomes were markedly similar, consistent with a single origin of the F508del allele. Together, these high-resolution variant analyses of the CFTR locus suggest a role for non-coding regulatory motifs in trait variation among individuals carrying the common CF allele. PMID:27917292

  16. Analytical methods to determine the comparative DNA binding studies of curcumin-Cu(II) complexes.

    Science.gov (United States)

    Rajesh, Jegathalaprathaban; Rajasekaran, Marichamy; Rajagopal, Gurusamy; Athappan, Periakaruppan

    2012-11-01

    DNA interaction studies of two mononuclear [1:1(1); 1:2(2)] copper(II) complexes of curcumin have been studied. The interaction of these complexes with CT-DNA has been explored by physical methods to propose modes of DNA binding of the complexes. Absorption spectral titrations of complex 1 with CT-DNA shows a red-shift of 3 nm with the DNA binding affinity of K(b), 5.21×10(4)M(-1) that are higher than that obtained for 2 (red-shift, 2 nm; K(b), 1.73×10(4)M(-1)) reveal that the binding occurs in grooves as a result of the interaction is via exterior phosphates. The CD spectra of these Cu(II) complexes show a red shift of 3-10nm in the positive band with increase in intensities. This spectral change of induced CD due to the hydrophobic interaction of copper complexes with DNA is the characteristic of B to A conformational change. The EB displacement assay also reveals the same trend as observed in UV-Vis spectral titration. The addition of complexes 1 and 2 to the DNA bound ethidium bromide (EB) solutions causes an obvious reduction in emission intensities indicating that these complexes competitively bind to DNA with EB. The positive shift of both the E(pc) and E(0)' accompanied by reduction of peak currents in differential pulse voltammogram (DPV), upon adding different concentrations of DNA to the metal complexes, are obviously in favor of strong binding to DNA. The super coiled plasmid pUC18 DNA cleavage ability of Cu(II) complexes in the presence of reducing agent reveals the single strand DNA cleavage (ssDNA) is observed. The hydroxyl radical (HO()) and the singlet oxygen are believed to be the reactive species responsible for the cleavage.

  17. Characterization of PCS1055, a novel muscarinic M4 receptor antagonist.

    Science.gov (United States)

    Croy, Carrie H; Chan, Wai Y; Castetter, Andrea M; Watt, Marla L; Quets, Anne T; Felder, Christian C

    2016-07-05

    Identification of synthetic ligands selective for muscarinic receptor subtypes has been challenging due to the high sequence identity and structural homology among the five muscarinic acetylcholine receptors. Here, we report the pharmacological characterization of PCS1055, a novel muscarinic M4 receptor antagonist. PCS1055 inhibited radioligand [(3)H]-NMS binding to the M4 receptor with a Ki=6.5nM. Though the potency of PCS1055 is lower than that of pan-muscarinic antagonist atropine, it has better subtype selectivity over previously reported M4-selective reagents such as the muscarinic-peptide toxins (Karlsson et al., 1994; Santiago and Potter, 2001a) at the M1 subtype, and benzoxazine ligand PD102807 at the M3-subtype (Bohme et al., 2002). A detailed head-to-head comparison study using [(3)H]-NMS competitive binding assays characterizes the selectivity profiles of PCS1055 to that of other potent muscarinic-antagonist compounds PD102807, tropicamide, AF-DX-384, pirenzapine, and atropine. In addition to binding studies, the subtype specificity of PCS1055 is also demonstrated by functional receptor activation as readout by GTP-γ-[(35)S] binding. These GTP-γ-[(35)S] binding studies showed that PCS1055 exhibited 255-, 69.1-, 342- and >1000-fold greater inhibition of Oxo-M activity at the M4 versus the M1-, M2(-), M3-or M5 receptor subtypes, respectively. Schild analyses indicates that PCS1055 acts as a competitive antagonist to muscarinic M4 receptor, and confirms the affinity of the ligand to be low nanomolar, Kb=5.72nM. Therefore, PCS1055 represents a new M4-preferring antagonist that may be useful in elucidating the roles of M4 receptor signaling.

  18. Genomic structure, organisation, and promoter analysis of the bovine (Bos taurus) Mx1 gene.

    Science.gov (United States)

    Gérardin, Joël A; Baise, Etienne A; Pire, Grégory A; Leroy, Michaël P-P; Desmecht, Daniel J-M

    2004-02-04

    Some MX proteins are known to confer a specific resistance against a panel of single-stranded RNA viruses. Many diseases due to such viruses are known to affect cattle worldwide, raising the possibility that the identification of an antiviral isoform of a bovine MX protein would allow the implementation of genetic selection programs aimed at improving innate resistance of cattle. With this potential application in mind, the present study was designed to isolate the bovine Mx1 gene including its promoter region and to investigate its genomic organisation and promoter reactivity. The bovine Mx1 gene is made up of 15 exons. All exon-intron boundaries conformed to the consensus sequences. A PCR product that contained a approximately 1-kb, 5'-flanking region upstream from the putative transcription start site was sequenced. Unexpectedly, this DNA region did not contain TATA or CCAAT motifs. A computer scan of the region disclosed a series of putative binding sites for known cytokines and transcription factors. There was a GAAAN(1-2)GAAA(C/G) motif, typical of an interferon-sensitive responsive element, between -118 and -107 from the putative transcription start site. There were also a NF-kappaB, two interleukin-6 binding sites, two Sp1 sites and five GC-rich boxes. The region also contained 12 stretches of the GAAA type, as described in all IFN-inducible genes. Bovine Mx1 expression was assessed by Northern blotting and immunofluorescence in the Madin Darby bovine kidney cells (MDBK) cell line treated with several stimuli. In conclusion, the bovine Mx1 gene and promoter region share the major structural and functional characteristics displayed by their homologs described in the rainbow trout, chicken, mouse and man.

  19. The Schizosaccharomyces pombe inv1+ regulatory region is unusually large and contains redundant cis-acting elements that function in a SAGA- and Swi/Snf-dependent fashion.

    Science.gov (United States)

    Ahn, Sejin; Spatt, Dan; Winston, Fred

    2012-08-01

    The Schizosaccharomyces pombe inv1(+) gene encodes invertase, the enzyme required for hydrolysis of sucrose and raffinose. Transcription of inv1(+) is regulated by glucose levels, with transcription tightly repressed in high glucose and strongly induced in low glucose. To understand this regulation, we have analyzed the inv1(+) cis-regulatory region and the requirement for the trans-acting coactivators SAGA and Swi/Snf. Surprisingly, deletion of the entire 1-kilobase intergenic region between the inv1(+) TATA element and the upstream open reading frame SPCC191.10 does not significantly alter regulation of inv1(+) transcription. However, a longer deletion that extends through SPCC191.10 abolishes inv1(+) induction in low glucose. Additional analysis demonstrates that there are multiple, redundant regulatory regions spread over 1.5 kb 5' of inv1(+), including within SPCC191.10, that can confer glucose-mediated transcriptional regulation to inv1(+). Furthermore, SPCC191.10 can regulate inv1(+) transcription in an orientation-independent fashion and from a distance as great as 3 kb. With respect to trans-acting factors, both SAGA and Swi/Snf are recruited to SPCC191.10 and to other locations in the large inv1(+) regulatory region in a glucose-dependent fashion, and both are required for inv1(+) derepression. Taken together, these results demonstrate that inv1(+) regulation in S. pombe occurs via the use of multiple regulatory elements and that activation can occur over a great distance, even from elements within other open reading frames.

  20. Extrachromosomal recombination in vaccinia-infected cells requires a functional DNA polymerase participating at a level other than DNA replication.

    Science.gov (United States)

    Colinas, R J; Condit, R C; Paoletti, E

    1990-12-01

    Homologous recombination was measured in vaccinia-infected cells cotransfected with two plasmid recombination substrates. One plasmid contains a vaccinia protein lacZ coding region bearing a 1.1 kb 3' terminal deletion while the other plasmid contains a non-promoted lacZ coding region bearing a 1.1 kb 5' terminal deletion. Homologous recombination occurring between the 825 bp of lacZ common to both plasmids regenerates a functional lacZ gene from which B-galactosidase expression was measured. The entire 3 kb lacZ gene was used as a positive control. A panel of thermosensitive mutants was screened in cells either transfected with the positive control plasmid or cotransfected with the recombination substrates. A DNA - mutant, ts42, known to map to the viral DNA polymerase gene was found to be defective in recombination. Significantly, other DNA - mutants, ts17 or ts25, or other DNA polymerase mutants did not exhibit a defect in recombination similar to ts42. Inhibitors of viral DNA synthesis did not uniformly affect recombination. Cytosine arabinoside and aphidicolin inhibited B-galactosidase expression from the recombination substrates but not from the positive control plasmid, whereas hydroxyurea enhanced expression from both. Marker rescue with the cloned wildtype DNA polymerase gene repaired the defect in ts42. Southern and western analyses demonstrated that B-galactosidase activity was consistent with a recombined lacZ gene and unit size 116 kDa protein. Measurement of plasmid and viral DNA replication in cells infected with the different DNA - mutants indicated that recombination was independent of plasmid and viral DNA replication. Together these results suggest that the vaccinia DNA polymerase participates in homologous recombination at a level other than that of DNA replication.

  1. The impact of cHS4 insulators on DNA transposon vector mobilization and silencing in retinal pigment epithelium cells.

    Directory of Open Access Journals (Sweden)

    Nynne Sharma

    Full Text Available DNA transposons have become important vectors for efficient non-viral integration of transgenes into genomic DNA. The Sleeping Beauty (SB, piggyBac (PB, and Tol2 transposable elements have distinct biological properties and currently represent the most promising transposon systems for animal transgenesis and gene therapy. A potential obstacle, however, for persistent function of integrating vectors is transcriptional repression of the element and its genetic cargo. In this study we analyze the insulating effect of the 1.2-kb 5'-HS4 chicken β-globin (cHS4 insulator element in the context of SB, PB, and Tol2 transposon vectors. By examining transgene expression from genomically inserted transposon vectors encoding a marker gene driven by a silencing-prone promoter, we detect variable levels of transcriptional silencing for the three transposon systems in retinal pigment epithelium cells. Notably, the PB system seems less vulnerable to silencing. Incorporation of cHS4 insulator sequences into the transposon vectors results in 2.2-fold and 1.5-fold increased transgene expression levels for insulated SB and PB vectors, respectively, but an improved persistency of expression was not obtained for insulated transgenes. Colony formation assays and quantitative excision assays unveil enhanced SB transposition efficiencies by the inclusion of the cHS4 element, resulting in a significant increase in the stable transfection rate for insulated SB transposon vectors in human cell lines. Our findings reveal a positive impact of cHS4 insulator inclusion for SB and PB vectors in terms of increased transgene expression levels and improved SB stable transfection rates, but also the lack of a long-term protective effect of the cHS4 insulator against progressive transgene silencing in retinal pigment epithelium cells.

  2. THAP and ATF-2 Regulated Sterol Carrier Protein-2 Promoter Activities in the Larval Midgut of the Yellow Fever Mosquito, Aedes aegypti

    Science.gov (United States)

    Peng, Rong; Fu, Qiang; Hong, Huazhu; Schwaegler, Tyler; Lan, Que

    2012-01-01

    Expression of sterol carrier protein-2 (SCP-2) in Aedes aegypti shows a distinct temporal/spatial pattern throughout the life cycle. In order to identify the transcription factors responsible for the larval temporal/spatial regulation of AeSCP-2 transcription, AeSCP-2 promoter activities were studied in vivo via transient transfection of promoter/reporter gene assays. Regulatory sequences upstream −1.3 kb of the transcription start site of AeSCP-2 were found to be critical for the in vivo temporal/spatial promoter activity. Interestingly, the −1.6 kb promoter sequence efficiently drove the larval midgut-specific siRNA expression, indicating that the −1.6 kb upstream sequence is sufficient for temporal/spatial AeSCP-2 transcriptional activity. Four transcription factors were identified in the midgut nuclear extract from feeding larvae via labeled −1.6/−1.3 kb DNA probe pull-down and proteomic analysis. Co-transfection of the promoter/reporter gene with inducible siRNA expression of each transcription factor was performed to confirm the regulatory function of individual transcription factor on AeSCP-2 transcriptional activities in the larval midgut. The results indicate that two of the identified transcription factors, Thanatos-associated protein (THAP) and activating transcription factor-2 (ATF-2), antagonistically control AeSCP-2 transcriptional activity in the midgut of feeding larvae via the regulatory sequences between −1.6 to −1.3 kb 5′ upstream of the transcription start site. In vivo expression knockdown of THAP and ATF-2 resulted in significant changes in developmental progression, which may be partially due to their effects on AeSCP-2 expression. PMID:23056538

  3. The 5'-flanking region of the RP58 coding sequence shows prominent promoter activity in multipolar cells in the subventricular zone during corticogenesis.

    Science.gov (United States)

    Ohtaka-Maruyama, C; Hirai, S; Miwa, A; Takahashi, A; Okado, H

    2012-01-10

    Pyramidal neurons of the neocortex are produced from progenitor cells located in the neocortical ventricular zone (VZ) and subventricular zone (SVZ) during embryogenesis. RP58 is a transcriptional repressor that is strongly expressed in the developing brain and plays an essential role in corticogenesis. The expression of RP58 is strictly regulated in a time-dependent and spatially restricted manner. It is maximally expressed in E15-16 embryonic cerebral cortex, localized specifically to the cortical plate and SVZ of the neocortex, hippocampus, and parts of amygdala during brain development, and found in glutamatergic but not GABAergic neurons. Identification of the promoter activity underlying specific expression patterns provides important clues to their mechanisms of action. Here, we show that the RP58 gene promoter is activated prominently in multipolar migrating cells, the first in vivo analysis of RP58 promoter activity in the brain. The 5.3 kb 5'-flanking genomic DNA of the RP58 coding region demonstrates promoter activity in neurons both in vitro and in vivo. This promoter is highly responsive to the transcription factor neurogenin2 (Ngn2), which is a direct upstream activator of RP58 expression. Using in utero electroporation, we demonstrate that RP58 gene promoter activity is first detected in a subpopulation of pin-like VZ cells, then prominently activated in migrating multipolar cells in the multipolar cell accumulation zone (MAZ) located just above the VZ. In dissociated primary cultured cortical neurons, RP58 promoter activity mimics in vivo expression patterns from a molecular standpoint that RP58 is expressed in a fraction of Sox2-positive progenitor cells, Ngn2-positive neuronal committed cells, and Tuj1-positive young neurons, but not in Dlx2-positive GABAergic neurons. Finally, we show that Cre recombinase expression under the control of the RP58 gene promoter is a feasible tool for conditional gene switching in post-mitotic multipolar migrating

  4. Molecular cloning and characterization of the human beta-like globin gene cluster.

    Science.gov (United States)

    Fritsch, E F; Lawn, R M; Maniatis, T

    1980-04-01

    The genes encoding human embryonic (epsilon), fetal (G gamma, A gamma) and adult (delta, beta) beta-like globin polypeptides were isolated as a set of overlapping cloned DNA fragments from bacteriophage lambda libraries of high molecular weight (15-20 kb) chromosomal DNA. The 65 kb of DNA represented in these overlapping clones contains the genes for all five beta-like polypeptides, including the embryonic epsilon-globin gene, for which the chromosomal location was previously unknown. All five genes are transcribed from the same DNA strand and are arranged in the order 5'-epsilon-(13.3 kb)-G gamma-(3.5 kb)-A gamma-(13.9 kb)-delta-(5.4 kb)-beta-3'. Thus the genes are positioned on the chromosome in the order of their expression during development. In addition to the five known beta-like globin genes, we have detected two other beta-like globin sequences which do not correspond to known polypeptides. One of these sequences has been mapped to the A gamma-delta intergenic region while the other is located 6-9 kb 5' to the epsilon gene. Cross hybridization experiments between the intergenic sequences of the gene cluster have revealed a nonglobin repeat sequence (*) which is interspersed with the globin genes in the following manner: 5'-**epsilon-*G gamma-A gamma*-**delta-beta*-3'. Fine structure mapping of the region located 5' to the delta-globin gene revealed two repeats with a maximum size of 400 bp, which are separated by approximately 700 bp of DNA not repeated within the cluster. Preliminary experiments indicate that this repeat family is also repeated many times in the human genome.

  5. Sequence variations in the 5' flanking and IVS-II regions of the G gamma- and A gamma-globin genes of beta S chromosomes with five different haplotypes.

    Science.gov (United States)

    Lanclos, K D; Oner, C; Dimovski, A J; Gu, Y C; Huisman, T H

    1991-06-01

    We have amplified and sequenced the 5' flanking and the second intervening sequence (IVS-II) regions of both the G gamma- and A gamma-globin genes of the beta S chromosomes from sickle cell anemia (SS) patients with homozygosities for five different haplotypes. The sequencing data, compared with previously published sequences for the normal chromosomes A and B, show many similarities to chromosome B for haplotypes 19, 20, and 17, while haplotypes 3 and 31 are remarkably similar to chromosome A and also similar to each other. Several unique mutations were found in the 5' flanking regions (G gamma and A gamma) of haplotypes 19 and 20 and in the IVS-II segments of the same genes of haplotypes 19, 20, and 17; the IVS-II of haplotypes 3 and 31 were identical to those of chromosome A. Dot-blot analyses of amplified DNA from additional SS patients with specific probes have confirmed that these mutations are unique for each haplotype. The two general patterns that have been observed among the five haplotypes have most probably arisen by gene conversion events between the A and B type chromosomes in the African population. These patterns correlate with high and low fetal hemoglobin expression, and it is speculated that these and other yet unknown gene conversions may contribute to the variations in hemoglobin F and G gamma levels observed among SS patients. In vitro expression experiments involving the approximately 1.3-kb 5' flanking regions of the G gamma- and A gamma-globin genes of the beta S chromosomes with the five different haplotypes failed to detect differences between the levels of expression, suggesting that the sequence variations observed between these segments of DNA are not the primary cause of the differences in hemoglobin F levels among the SS patients.

  6. Visual Performance and Optical Quality of Standardized Asymmetric Soft Contact Lenses in Patients With Keratoconus.

    Science.gov (United States)

    Suzaki, Asaki; Maeda, Naoyuki; Fuchihata, Mutsumi; Koh, Shizuka; Nishida, Kohji; Fujikado, Takashi

    2017-06-01

    To evaluate the visual performance and optical quality of a standardized asymmetric soft contact lens (SCL) used for correction of higher-order aberrations (HOAs) in eyes with keratoconus. We included 30 eyes (26 patients) with keratoconus (average K: 45.7 ± 2.3 diopters [D]). The patients were subjected to corneal tomography, aberrometry, measurements of manifest refraction and visual acuity (VA), and visual analog scale (VAS) assessments. The study lenses were made using a molding method and consisted of six standardized types, in which an asymmetric power distribution of approximately 2 to 12 D (2-D step) was used to correct HOAs. The lens type suitable for each eye was selected based on the corneal tomography and aberrometry data. The on-eye performance of the lens was evaluated using aberrometry (4-mm pupil), over refraction, VA, and VAS. The standardized asymmetric SCL improved the best spectacle-corrected VA from -0.07 ± 0.09 to -0.11 ± 0.08 logMAR (P keratoconus who wear rigid gas-permeable lenses.

  7. The surface temperature of Europa

    CERN Document Server

    Ashkenazy, Yosef

    2016-01-01

    Previous estimates of the surface temperature of Jupiter's moon, Europa, neglected the effect of the eccentricity of Jupiter's orbit around the Sun, the effect of the eclipse of Europa (i.e., the relative time that Europa is within the shadow of Jupiter), and the effect of Europa's internal heating. Here we estimate the surface temperature of Europa, when Europa's obliquity, eclipse and internal heating, as well as the eccentricity of Jupiter, are all taken into account. For a typical internal heating rate of 0.05 W/m$^2$ (corresponding to an ice thickness of about 10 kms), the equator, pole, and global mean surface temperatures are 101.7 K, 45.26 K, and 94.75 K, respectively. We found that the temperature at the high latitudes is significantly affected by the internal heating. We also studied the effect of the internal heating on the mean thickness of Europa's icy shell and conclude that the polar region temperature can be used to constrain the internal heating and the depth of the ice. Our approach and form...

  8. Effect of Backing Plate Thermal Property on Friction Stir Welding of 25-mm-Thick AA6061

    Science.gov (United States)

    Upadhyay, Piyush; Reynolds, Anthony

    2014-04-01

    By using backing plates made out of materials with widely varying thermal diffusivity this work seeks to elucidate the effects of the root side thermal boundary condition on weld process variables and resulting joint properties. Welds were made in 25.4-mm-thick AA6061 using ceramic, titanium, steel, and aluminum as backing plate (BP) material. Welds were also made using a "composite backing plate" consisting of longitudinal narrow strip of low diffusivity material at the center and two side plates of high diffusivity aluminum. Stir zone temperature during the welding was measured using two thermocouples spot welded at the core of the probe: one at the midplane height and another near the tip of the probe corresponding to the root of the weld. Steady state midplane probe temperatures for all the BPs used were found to be very similar. Near root peak temperature, however, varied significantly among weld made with different BPs all other things being equal. Whereas the near root and midplane temperature were the same in the case of ceramic backing plate, the root peak temperature was 318 K (45 °C) less than the midplane temperature in the case of aluminum BP. The trends of nugget hardness and grain size in through thickness direction were in agreement with the measured probe temperatures. Hardness and tensile test results show that the use of composite BP results in stronger joint compared to monolithic steel BP.

  9. High Performance InAs/In0.53Ga0.23Al0.24As/InP Quantum Dot 1.55 um Tunnel Injection Laser

    KAUST Repository

    Bhowmick, Sishir

    2014-01-01

    The characteristics of 1.55 ? InAs self-organized quantum-dot lasers, grown on (001) InP substrates by molecular beam epitaxy, have been investigated. Modulation doping of the dots with holes and tunnel injection of electrons have been incorporated in the design of the active (gain) region of the laser heterostructure. Large values of To=227 K (5 °C ? T ?45 °C) and 100 K (45 °C ? T ? 75 °C) were derived from temperature dependent measurements of the light-current characteristics. The modal gain per dot layer is 14.5 cm -1 and the differential gain derived from both light-current and small-signal modulation measurements is 0.8}\\times 10-15 cm}2. The maximum measured 3 rm dB small-signal modulation bandwidth is 14.4 GHz and the gain compression factor is 5.4\\times 10-17 cm}2. The lasers are characterized by a chirp of 0.6 AA for a modulation frequency of 10 GHz and a near zero ?-parameter at the peak of the laser emission. These characteristics are amongst the best from any 1.55 ? edge-emitting semiconductor laser. © 1965-2012 IEEE.

  10. Hypochlorous acid-mediated oxidation of lipid components and antioxidants present in low-density lipoproteins

    DEFF Research Database (Denmark)

    Pattison, David I; Hawkins, Clare Louise; Davies, Michael Jonathan

    2003-01-01

    and antioxidants in aqueous solution (pH 7.4). The reactions of HOCl with phosphoryl-serine and phosphoryl-ethanolamine are rapid (k approximately 10(5) M(-)(1) s(-)(1)) and of comparable reactivity to many of the protein sites. The major products formed in these reactions are chloramines, which decay to give both...... and yielded k = 9 M(-)(1) s(-)(1). The reactions of the lipid-soluble antioxidants, alpha-tocopherol and ubiquinol-10, with HOCl were investigated with model compounds. For the reactions of HOCl with both Trolox and ubiquinol-0, k = 1.3 x 10(3) M(-)(1) s(-)(1); thus, these lipid soluble antioxidants...... are relatively ineffective as direct scavengers for HOCl as compared to water soluble antioxidants (e.g., ascorbate, k ca. 10(6) M(-)(1) s(-)(1)). The reaction of HOCl with hydroquinone (a simple model for ubiquinol-10) was also investigated both in aqueous solution (k = 45 M(-)(1) s(-)(1)) and in a less polar...

  11. Structure-dependent charge density as a determinant of antimicrobial activity of peptide analogues of defensin.

    Science.gov (United States)

    Bai, Yang; Liu, Shouping; Jiang, Ping; Zhou, Lei; Li, Jing; Tang, Charles; Verma, Chandra; Mu, Yuguang; Beuerman, Roger W; Pervushin, Konstantin

    2009-08-01

    Defensins are small (3-5 kDa) cysteine-rich cationic proteins found in both vertebrates and invertebrates constituting the front line of host innate immunity. Despite intensive research, bactericidal and cytotoxic mechanisms of defensins are still largely unknown. Moreover, we recently demonstrated that small peptides derived from defensins are even more potent bactericidal agents with less toxicity toward host cells. In this paper, structures of three C-terminal (R36-K45) analogues of human beta-defensin-3 were studied by 1H NMR spectroscopy and extensive molecular dynamics simulations. Because of indications that these peptides might target the inner bacterial membrane, they were reconstituted in dodecylphosphocholine or dodecylphosphocholine/1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] mixed micelles, and lipid bicelles mimicking the phospholipid-constituted bilayer membrane of mammalian and bacterial cells. The results show that the binding affinity and partitioning into the lipid phase and the ability to dimerize and accrete well-defined structures upon interactions with lipid membranes contribute to compactization of positive charges within peptide oligomers. The peptide charge density, mediated by corresponding three-dimensional structures, was found to directly correlate with the antimicrobial activity. These novel observations may provide a new rationale for the design of improved antimicrobial agents.

  12. Efficient transformation of the yellow fever mosquito Aedes aegypti using the piggyBac transposable element vector pBac[3xP3-EGFP afm].

    Science.gov (United States)

    Kokoza, V; Ahmed, A; Wimmer, E A; Raikhel, A S

    2001-11-01

    We report efficient germ-line transformation in the yellow fever mosquito Aedes aegypti accomplished using the piggyBac transposable element vector pBac[3xP3-EGFP afm]. Two transgenic lines were established and characterized; each contained the Vg-Defensin A transgene with strong eye-specific expression of the enhanced green fluorescent protein (EGFP) marker gene regulated by the artificial 3xP3 promoter. Southern blot hybridization and inverse PCR analyses of genomic DNA demonstrated a precise piggyBac-mediated, single copy insertion of the pBac[3xP3-EGFP afm,Vg-DefA] transposon in each transgenic line. For each line, genetic analysis confirmed stability and integrity of the entire transposon construct in the mosquito genome through the G2-G6 generations. Successful establishment of homozygous transgenic lines indicated that in both cases a non-lethal integration of the transposon into the mosquito genome had occurred. The 3xP3-EGFP marker was tested in mosquitoes with different genetic backgrounds. In white-eyed transgenic mosquitoes, the strong eye-specific expression of GFP was observed throughout all stages of development, starting from newly hatched first instar larvae to adults. A similar level and pattern of fluorescence was observed in red-eyed mosquitoes that were generated by crossing the 3xP3-EGFP transformants with the kh(w) white-eye mosquitoes transformed with the Drosophila cinnabar gene. Importantly, the utility of the 3xP3-EGFP, as marker gene for transformation of wild type mosquitoes, was demonstrated by strong eye-specific GFP expression in larval and pupal stages of black-eyed hybrids of the 3xP3-EGFP white-eye transformants and the wild type Rockefeller/UGAL strain. Finally, analysis of the Vg-DefA transgene expression in transformants from two established lines demonstrated strong blood-meal activation and fat-body-specific expression regulated by the Vg 1.8-kb 5' upstream region.

  13. Identification of novel androgen receptor target genes in prostate cancer

    Directory of Open Access Journals (Sweden)

    Gerald William L

    2007-06-01

    Full Text Available Abstract Background The androgen receptor (AR plays critical roles in both androgen-dependent and castrate-resistant prostate cancer (PCa. However, little is known about AR target genes that mediate the receptor's roles in disease progression. Results Using Chromatin Immunoprecipitation (ChIP Display, we discovered 19 novel loci occupied by the AR in castrate resistant C4-2B PCa cells. Only four of the 19 AR-occupied regions were within 10-kb 5'-flanking regulatory sequences. Three were located up to 4-kb 3' of the nearest gene, eight were intragenic and four were in gene deserts. Whereas the AR occupied the same loci in C4-2B (castrate resistant and LNCaP (androgen-dependent PCa cells, differences between the two cell lines were observed in the response of nearby genes to androgens. Among the genes strongly stimulated by DHT in C4-2B cells – D-dopachrome tautomerase (DDT, Protein kinase C delta (PRKCD, Glutathione S- transferase theta 2 (GSTT2, Transient receptor potential cation channel subfamily V member 3 (TRPV3, and Pyrroline-5-carboxylate reductase 1 (PYCR1 – most were less strongly or hardly stimulated in LNCaP cells. Another AR target gene, ornithine aminotransferase (OAT, was AR-stimulated in a ligand-independent manner, since it was repressed by AR siRNA knockdown, but not stimulated by DHT. We also present evidence for in vivo AR-mediated regulation of several genes identified by ChIP Display. For example, PRKCD and PYCR1, which may contribute to PCa cell growth and survival, are expressed in PCa biopsies from primary tumors before and after ablation and in metastatic lesions in a manner consistent with AR-mediated stimulation. Conclusion AR genomic occupancy is similar between LNCaP and C4-2B cells and is not biased towards 5' gene flanking sequences. The AR transcriptionally regulates less than half the genes nearby AR-occupied regions, usually but not always, in a ligand-dependent manner. Most are stimulated and a few are

  14. The Juan non-LTR retrotransposon in mosquitoes: genomic impact, vertical transmission and indications of recent and widespread activity

    Directory of Open Access Journals (Sweden)

    Tu Zhijian

    2007-07-01

    Full Text Available Abstract Background In contrast to DNA-mediated transposable elements (TEs, retrotransposons, particularly non-long terminal repeat retrotransposons (non-LTRs, are generally considered to have a much lower propensity towards horizontal transfer. Detailed studies on site-specific non-LTR families have demonstrated strict vertical transmission. More studies are needed with non-site-specific non-LTR families to determine whether strict vertical transmission is a phenomenon related to site specificity or a more general characteristic of all non-LTRs. Juan is a Jockey clade non-LTR retrotransposon first discovered in mosquitoes that is widely distributed in the mosquito family Culicidae. Being a non-site specific non-LTR, Juan offers an opportunity to further investigate the hypothesis that non-LTRs are genomic elements that are primarily vertically transmitted. Results Systematic analysis of the ~1.3 Gbp Aedes aegypti (Ae. aegypti genome sequence suggests that Juan-A is the only Juan-type non-LTR in Aedes aegypti. Juan-A is highly reiterated and comprises approximately 3% of the genome. Using minimum cutoffs of 90% length and 70% nucleotide (nt identity, 663 copies were found by BLAST using the published Juan-A sequence as the query. All 663 copies are at least 95% identical to Juan-A, while 378 of these copies are 99% identical to Juan-A, indicating that the Juan-A family has been transposing recently in evolutionary history. Using the 0.34 Kb 5' UTR as the query, over 2000 copies were identified that may contain internal promoters, leading to questions on the genomic impact of Juan-A. Juan sequences were obtained by PCR, library screening, and database searches for 18 mosquito species of six genera including Aedes, Ochlerotatus, Psorophora, Culex, Deinocerites, and Wyeomyia. Comparison of host and Juan phylogenies shows overall congruence with few exceptions. Conclusion Juan-A is a major genomic component in Ae. aegypti and it has been

  15. An X-linked homologue of the autosomal inprinted gene ZNF127 escapes X inactivation

    Energy Technology Data Exchange (ETDEWEB)

    Longstreet, M.; Nicholls, R.D.; Willard, H.F. [Case Western Reserve Univ., Cleveland, OH (United States)] [and others

    1994-09-01

    The ZNF127 gene has been shown to be subject to parental imprinting in both humans and the mouse and maps to within the Prader-Willi/Angelman Syndrome critical region on chromosome 15. We have cloned two X-linked related loci, one of which, ZNFXp is a transcribed gene while the other, ZNFXq, is an untranscribed pseudogene. ZNFXp is 83.6% identical to ZNFXq and 65.4% identical to ZNF127 over 1.4 kb of open reading frame they share in common, Like ZNF127, the predicted protein sequence of ZNFXp contains a C{sub 3}HC{sub 4} zinc finger domain and C{sub 3}H zinc finger-like motifs. Whereas ZNF127 has three C{sub 3}H motifs, ZNFXp has four. A strong CpG island is located within 1 kb 5{prime} of the predicted amino terminus of ZNFXp. Expression of ZNFXp has been detected from mouse/human somatic cell hybrids containing either an active (n=2) or an inactive (n=4) chromosome, and thus escapes X inactivation. Probes made from the 3{prime} UTR of ZNFXp detect a number of related loci in both human and murine DNA, none of which is the ZNF127 locus on chromosome 15. None of the detectable murine bands shows dosage differences between males and females as would be expected for X-linked loci. This raises the possibility that ZNFXp inserted into the human X chromosome after its divergence from a common ancestor with the murine X. We have mapped ZNFXp to Xp11.4 by Southern blotting and PCR of hybrid DNAs and by fluorescence in situ hybridization (FISH). ZNFXq maps within the X Inactivation Center (XIC) region on Xq13.2, approximately 300 kb distal to the XIST gene. We find it intriguing, and perhaps significant, that two members of this gene family are subject to epigenetic regulation -- one autosomal imprinting, and the other escape from X inactivation. These results could imply an evolutionary and mechanistic relationship between these two processes.

  16. Spectroscopic studies of the binding of Cu(II) complexes of oxicam NSAIDs to alternating G-C and homopolymeric G-C sequences.

    Science.gov (United States)

    Chakraborty, Sreeja; Bose, Madhuparna; Sarkar, Munna

    2014-03-25

    Drugs belonging to the Non-steroidal anti-inflammatory (NSAID) group are not only used as anti-inflammatory, analgesic and anti-pyretic agents, but also show anti-cancer effects. Complexing them with a bioactive metal like copper, show an enhancement in their anti-cancer effects compared to the bare drugs, whose exact mechanism of action is not yet fully understood. For the first time, it was shown by our group that Cu(II)-NSAIDs can directly bind to the DNA backbone. The ability of the copper complexes of NSAIDs namely meloxicam and piroxicam to bind to the DNA backbone could be a possible molecular mechanism behind their enhanced anticancer effects. Elucidating base sequence specific interaction of Cu(II)-NSAIDs to the DNA will provide information on their possible binding sites in the genome sequence. In this work, we present how these complexes respond to differences in structure and hydration pattern of GC rich sequences. For this, binding studies of Cu(II) complexes of piroxicam [Cu(II)-(Px)2 (L)2] and meloxicam [Cu(II)-(Mx)2 (L)] with alternating GC (polydG-dC) and homopolymeric GC (polydG-polydC) sequences were carried out using a combination of spectroscopic techniques that include UV-Vis absorption, fluorescence and circular dichroism (CD) spectroscopy. The Cu(II)-NSAIDs show strong binding affinity to both polydG-dC and polydG-polydC. The role reversal of Cu(II)-meloxicam from a strong binder of polydG-dC (Kb=11.5×10(3) M(-1)) to a weak binder of polydG-polydC (Kb=5.02×10(3) M(-1)), while Cu(II)-piroxicam changes from a strong binder of polydG-polydC (Kb=8.18×10(3) M(-1)) to a weak one of polydG-dC (Kb=2.18×10(3) M(-1)), point to the sensitivity of these complexes to changes in the backbone structures/hydration. Changes in the profiles of UV absorption band and CD difference spectra, upon complex binding to polynucleotides and the results of competitive binding assay using ethidium bromide (EtBr) fluorescence indicate different binding modes in each

  17. Spectroscopic studies of the binding of Cu(II) complexes of oxicam NSAIDs to alternating G-C and homopolymeric G-C sequences

    Science.gov (United States)

    Chakraborty, Sreeja; Bose, Madhuparna; Sarkar, Munna

    2014-03-01

    Drugs belonging to the Non-steroidal anti-inflammatory (NSAID) group are not only used as anti-inflammatory, analgesic and anti-pyretic agents, but also show anti-cancer effects. Complexing them with a bioactive metal like copper, show an enhancement in their anti-cancer effects compared to the bare drugs, whose exact mechanism of action is not yet fully understood. For the first time, it was shown by our group that Cu(II)-NSAIDs can directly bind to the DNA backbone. The ability of the copper complexes of NSAIDs namely meloxicam and piroxicam to bind to the DNA backbone could be a possible molecular mechanism behind their enhanced anticancer effects. Elucidating base sequence specific interaction of Cu(II)-NSAIDs to the DNA will provide information on their possible binding sites in the genome sequence. In this work, we present how these complexes respond to differences in structure and hydration pattern of GC rich sequences. For this, binding studies of Cu(II) complexes of piroxicam [Cu(II)-(Px)2 (L)2] and meloxicam [Cu(II)-(Mx)2 (L)] with alternating GC (polydG-dC) and homopolymeric GC (polydG-polydC) sequences were carried out using a combination of spectroscopic techniques that include UV-Vis absorption, fluorescence and circular dichroism (CD) spectroscopy. The Cu(II)-NSAIDs show strong binding affinity to both polydG-dC and polydG-polydC. The role reversal of Cu(II)-meloxicam from a strong binder of polydG-dC (Kb = 11.5 × 103 M-1) to a weak binder of polydG-polydC (Kb = 5.02 × 103 M-1), while Cu(II)-piroxicam changes from a strong binder of polydG-polydC (Kb = 8.18 × 103 M-1) to a weak one of polydG-dC (Kb = 2.18 × 103 M-1), point to the sensitivity of these complexes to changes in the backbone structures/hydration. Changes in the profiles of UV absorption band and CD difference spectra, upon complex binding to polynucleotides and the results of competitive binding assay using ethidium bromide (EtBr) fluorescence indicate different binding modes in each

  18. Cloning and functional characterization of the 5' regulatory region of ovine Hormone Sensitive Lipase (HSL) gene.

    Science.gov (United States)

    Lampidonis, Antonis D; Stravopodis, Dimitrios J; Voutsinas, Gerassimos E; Messini-Nikolaki, Niki; Stefos, George C; Margaritis, Lukas H; Argyrokastritis, Alexandros; Bizelis, Iosif; Rogdakis, Emmanuel

    2008-12-31

    transiently transfected into 3T3-L1 (mouse fibroblasts) as well as T24 (human bladder cancer) cell lines, strong promoter activities were unambiguously detected, with the -140/+18 nucleotide sequence bearing the highest transcriptional response, thus indicating that the 1.224 kb 5' flanking region, isolated by genome walking, veritably contains the ovHSL gene promoter. Of particular significance are the observations that the functional promoter fragments could trigger the transcriptional activity of luciferase gene only under high concentration of glucose conditions in both cell lines.

  19. Asymmetrical distribution of non-conserved regulatory sequences at PHOX2B is reflected at the ENCODE loci and illuminates a possible genome-wide trend

    Directory of Open Access Journals (Sweden)

    McCallion Andrew S

    2009-01-01

    Full Text Available Abstract Background Transcriptional regulatory elements are central to development and interspecific phenotypic variation. Current regulatory element prediction tools rely heavily upon conservation for prediction of putative elements. Recent in vitro observations from the ENCODE project combined with in vivo analyses at the zebrafish phox2b locus suggests that a significant fraction of regulatory elements may fall below commonly applied metrics of conservation. We propose to explore these observations in vivo at the human PHOX2B locus, and also evaluate the potential evidence for genome-wide applicability of these observations through a novel analysis of extant data. Results Transposon-based transgenic analysis utilizing a tiling path proximal to human PHOX2B in zebrafish recapitulates the observations at the zebrafish phox2b locus of both conserved and non-conserved regulatory elements. Analysis of human sequences conserved with previously identified zebrafish phox2b regulatory elements demonstrates that the orthologous sequences exhibit overlapping regulatory control. Additionally, analysis of non-conserved sequences scattered over 135 kb 5' to PHOX2B, provides evidence of non-conserved regulatory elements positively biased with close proximity to the gene. Furthermore, we provide a novel analysis of data from the ENCODE project, finding a non-uniform distribution of regulatory elements consistent with our in vivo observations at PHOX2B. These observations remain largely unchanged when one accounts for the sequence repeat content of the assayed intervals, when the intervals are sub-classified by biological role (developmental versus non-developmental, or by gene density (gene desert versus non-gene desert. Conclusion While regulatory elements frequently display evidence of evolutionary conservation, a fraction appears to be undetected by current metrics of conservation. In vivo observations at the PHOX2B locus, supported by our analyses of in

  20. An Sp185/333 gene cluster from the purple sea urchin and putative microsatellite-mediated gene diversification

    Directory of Open Access Journals (Sweden)

    Buckley Katherine M

    2010-10-01

    Full Text Available Abstract Background The immune system of the purple sea urchin, Strongylocentrotus purpuratus, is complex and sophisticated. An important component of sea urchin immunity is the Sp185/333 gene family, which is significantly upregulated in immunologically challenged animals. The Sp185/333 genes are less than 2 kb with two exons and are members of a large diverse family composed of greater than 40 genes. The S. purpuratus genome assembly, however, contains only six Sp185/333 genes. This underrepresentation could be due to the difficulties that large gene families present in shotgun assembly, where multiple similar genes can be collapsed into a single consensus gene. Results To understand the genomic organization of the Sp185/333 gene family, a BAC insert containing Sp185/333 genes was assembled, with careful attention to avoiding artifacts resulting from collapse or artificial duplication/expansion of very similar genes. Twelve candidate BAC assemblies were generated with varying parameters and the optimal assembly was identified by PCR, restriction digests, and subclone sequencing. The validated assembly contained six Sp185/333 genes that were clustered in a 34 kb region at one end of the BAC with five of the six genes tightly clustered within 20 kb. The Sp185/333 genes in this cluster were no more similar to each other than to previously sequenced Sp185/333 genes isolated from three different animals. This was unexpected given their proximity and putative effects of gene homogenization in closely linked, similar genes. All six genes displayed significant similarity including both 5' and 3' flanking regions, which were bounded by microsatellites. Three of the Sp185/333 genes and their flanking regions were tandemly duplicated such that each repeated segment consisted of a gene plus 0.7 kb 5' and 2.4 kb 3' of the gene (4.5 kb total. Both edges of the segmental duplications were bounded by different microsatellites. Conclusions The high sequence

  1. Regulation of zebrafish CYP3A65 transcription by AHR2

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Chin-Teng; Chung, Hsin-Yu; Su, Hsiao-Ting; Tseng, Hua-Pin [Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan (China); Tzou, Wen-Shyong [Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan (China); Center of Excellence for Marine Bioenvironment and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan (China); Hu, Chin-Hwa, E-mail: chhu@mail.ntou.edu.tw [Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan (China); Center of Excellence for Marine Bioenvironment and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan (China)

    2013-07-15

    CYP3A proteins are the most abundant CYPs in the liver and intestines, and they play a pivotal role in drug metabolism. In mammals, CYP3A genes are induced by various xenobiotics through processes mediated by PXR. We previously identified zebrafish CYP3A65 as a CYP3A ortholog that is constitutively expressed in gastrointestinal tissues, and is upregulated by treatment with dexamethasone, rifampicin or tetrachlorodibenzo-p-dioxin (TCDD). However, the underlying mechanism of TCDD-mediated CYP3A65 transcription is unclear. Here we generated two transgenic zebrafish, Tg(CYP3A65S:EGFP) and Tg(CYP3A65L:EGFP), which contain 2.1 and 5.4 kb 5′ flanking sequences, respectively, of the CYP3A65 gene upstream of EGFP. Both transgenic lines express EGFP in larval gastrointestinal tissues in a pattern similar to that of the endogenous CYP3A65 gene. Moreover, EGFP expression can be significantly induced by TCDD exposure during the larval stage. In addition, EGFP expression can be stimulated by kynurenine, a putative AHR ligand produced during tryptophan metabolism. AHRE elements in the upstream regulatory region of the CYP3A65 gene are indispensible for basal and TCDD-induced transcription. Furthermore, the AHR2 DNA and ligand-binding domains are required to mediate effective CYP3A65 transcription. AHRE sequences are present in the promoters of many teleost CYP3 genes, but not of mammalian CYP3 genes, suggesting that AHR/AHR2-mediated transcription is likely a common regulatory mechanism for teleost CYP3 genes. It may also reflect the different environments that terrestrial and aquatic organisms encounter. - Highlights: • Tg(CYP3A65:EGFP) and CYP3A65 exhibits identical expression pattern. • CYP3A65 can be significantly induced by TCDD or kynurenine. • The AHRE elements are required to mediate CYP3A65 transcription. • The AHR2 DNA and ligand-binding domains are required for CYP3A65 transcription. • AHRE elements are present in many teleost CYP3 genes, but not in

  2. Common gene variants in the tumor necrosis factor (TNF and TNF receptor superfamilies and NF-kB transcription factors and non-Hodgkin lymphoma risk.

    Directory of Open Access Journals (Sweden)

    Sophia S Wang

    Full Text Available BACKGROUND: A promoter polymorphism in the pro-inflammatory cytokine tumor necrosis factor (TNF (TNF G-308A is associated with increased non-Hodgkin lymphoma (NHL risk. The protein product, TNF-alpha, activates the nuclear factor kappa beta (NF-kappaB transcription factor, and is critical for inflammatory and apoptotic responses in cancer progression. We hypothesized that the TNF and NF-kappaB pathways are important for NHL and that gene variations across the pathways may alter NHL risk. METHODOLOGY/PRINCIPAL FINDINGS: We genotyped 500 tag single nucleotide polymorphisms (SNPs from 48 candidate gene regions (defined as 20 kb 5', 10 kb 3' in the TNF and TNF receptor superfamilies and the NF-kappaB and related transcription factors, in 1946 NHL cases and 1808 controls pooled from three independent population-based case-control studies. We obtained a gene region-level summary of association by computing the minimum p-value ("minP test". We used logistic regression to compute odds ratios and 95% confidence intervals for NHL and four major NHL subtypes in relation to SNP genotypes and haplotypes. For NHL, the tail strength statistic supported an overall relationship between the TNF/NF-kappaB pathway and NHL (p = 0.02. We confirmed the association between TNF/LTA on chromosome 6p21.3 with NHL and found the LTA rs2844484 SNP most significantly and specifically associated with the major subtype, diffuse large B-cell lymphoma (DLBCL (p-trend = 0.001. We also implicated for the first time, variants in NFKBIL1 on chromosome 6p21.3, associated with NHL. Other gene regions identified as statistically significantly associated with NHL included FAS, IRF4, TNFSF13B, TANK, TNFSF7 and TNFRSF13C. Accordingly, the single most significant SNPs associated with NHL were FAS rs4934436 (p-trend = 0.0024, IRF4 rs12211228 (p-trend = 0.0026, TNFSF13B rs2582869 (p-trend = 0.0055, TANK rs1921310 (p-trend = 0.0025, TNFSF7 rs16994592 (p-trend = 0.0024, and TNFRSF13C rs6002551

  3. Molecular Cloning and Characterization of Human Homeobox Gene Nkx3.1 Promoter

    Institute of Scientific and Technical Information of China (English)

    An-LiJIANG; Jian-YeZHANG; CharlesYOUNG; Xiao-YanHU; Yong-MeiWANG; Zhi-FangLIU; Mei-LanHAO

    2004-01-01

    Nkx3.1 is a prostate-specific homeobox gene related strongly to prostate development and prostate cancer. To study its regulation of transcription, 1.06 kb 5′ flanking region of Nkx3.1 gene and its 5′ deletion mutants (861,617,417 and 238 bp) were obtained by PCR and cloned into pGL3-basic, a promoter-less luciferase reporter vector, to examine their promoter activities driving the reporter gene transcription, pRL-TK, a Renilla luciferase reporter vector was used as internal control, and pGL3-control and pGL3-basic were used as positive and negative control respectively. The promoter activities were determined by dual-luciferase reporter assay 48h after pGL3 constructs were cotransfected with pRL-TK into prostate cancer cell LNCaP. The results showed that dual-luciferase reporter assay (M/M2) of pGL3-1.06kb cotransfection with pRL-TK was 2.7, which was about 1.5-fold higher than that of pGL3-control cotransfection with pRL-TK and 50-fold higher than that of pGL3-basic cotransfection with pRL-TK. The results also showed that the relative activities (M1/M2) were 0.71, 0.84, 0.44 and 2.07 respectively for pGL3-861bp, pGL3-617bp, pGL3-417bp, pGL3-238bp, the last one still had 80% promoter activity compared with pGL3-1.06kb, which showed that deletion from 1.06kb to 238 bp had small effects on promoter activity. The conclusion was that the 238bp fragment containing a TATA box and two CAAT boxes had strong promoter activity. However, the deletion from 1.06kb to 861bp reduced activity 3.8-fold while the deletion from 417bp to 238bp enhanced activity 4.7-fold, which indicated that these deleted sequences might contain some important positive or negative regulatory elements. It will be important to identify the elements within the Nkx3.1 promoter that contribute to regulation of the gene transcription in the future studies.

  4. Molecular Cloning and Characterization of Human Homeobox Gene Nkx3.1 Promoter

    Institute of Scientific and Technical Information of China (English)

    An-Li JIANG; Jian-Ye ZHANG; Charles YOUNG; Xiao-Yan HU; Yong-Mei WANG; Zhi-Fang LIU; Mei-Lan HAO

    2004-01-01

    Nkx3.1 is a prostate-specific homeobox gene related strongly to prostate development andprostate cancer. To study its regulation of transcription, 1.06 kb 5 ′ flanking region of Nkx3.1 gene and its5 ′deletion mutants (861,617, 417 and 238 bp) were obtained by PCR and cloned into pGL3-basic, apromoter-less luciferase reporter vector, to examine their promoter activities driving the reporter genetranscription, pRL-TK, a Renilla luciferase reporter vector was used as internal control, and pGL3-controland pGL3-basic were used as positive and negative control respectively. The promoter activities were deter-mined by dual-luciferase reporter assay 48 h after pGL3 constructs were cotransfected with pRL-TK intoprostate cancer cell LNCaP. The results showed that dual-luciferase reporter assay (M1/M2) of pGL3-1.06 kbcotransfection with pRL-TK was 2.7, which was about 1.5-fold higher than that of pGL3-control cotrans-fection with pRL-TK and 50-fold higher than that of pGL3-basic cotransfection with pRL-TK. The resultsalso showed that the relative activities (M1/M2) were 0.71, 0.84, 0.44 and 2.07 respectively for pGL3-861 bp,pGL3-617 bp, pGL3-417 bp, pGL3-238 bp, the last one still had 80% promoter activity compared with pGL3-1.06 kb, which showed that deletion from 1.06 kb to 238 bp had small effects on promoter activity. Theconclusion was that the 238 bp fragment containing a TATA box and two CAAT boxes had strong promoteractivity. However, the deletion from 1.06 kb to 861 bp reduced activity 3.8-fold while the deletion from 417bp to 238 bp enhanced activity 4.7-fold, which indicated that these deleted sequences might contain someimportant positive or negative regulatory elements. It will be important to identify the elements within theNkx3.1 promoter that contribute to regulation of the gene transcription in the future studies.

  5. RIP1 suppresses innate immune necrotic as well as apoptotic cell death during mammalian parturition.

    Science.gov (United States)

    Kaiser, William J; Daley-Bauer, Lisa P; Thapa, Roshan J; Mandal, Pratyusha; Berger, Scott B; Huang, Chunzi; Sundararajan, Aarthi; Guo, Hongyan; Roback, Linda; Speck, Samuel H; Bertin, John; Gough, Peter J; Balachandran, Siddharth; Mocarski, Edward S

    2014-05-27

    The pronecrotic kinase, receptor interacting protein (RIP1, also called RIPK1) mediates programmed necrosis and, together with its partner, RIP3 (RIPK3), drives midgestational death of caspase 8 (Casp8)-deficient embryos. RIP1 controls a second vital step in mammalian development immediately after birth, the mechanism of which remains unresolved. Rip1(-/-) mice display perinatal lethality, accompanied by gross immune system abnormalities. Here we show that RIP1 K45A (kinase dead) knockin mice develop normally into adulthood, indicating that development does not require RIP1 kinase activity. In the face of complete RIP1 deficiency, cells develop sensitivity to RIP3-mixed lineage kinase domain-like-mediated necroptosis as well as to Casp8-mediated apoptosis activated by diverse innate immune stimuli (e.g., TNF, IFN, double-stranded RNA). When either RIP3 or Casp8 is disrupted in combination with RIP1, the resulting double knockout mice exhibit slightly prolonged survival over RIP1-deficient animals. Surprisingly, triple knockout mice with combined RIP1, RIP3, and Casp8 deficiency develop into viable and fertile adults, with the capacity to produce normal levels of myeloid and lymphoid lineage cells. Despite the combined deficiency, these mice sustain a functional immune system that responds robustly to viral challenge. A single allele of Rip3 is tolerated in Rip1(-/-)Casp8(-/-)Rip3(+/-) mice, contrasting the need to eliminate both alleles of either Rip1 or Rip3 to rescue midgestational death of Casp8-deficient mice. These observations reveal a vital kinase-independent role for RIP1 in preventing pronecrotic as well as proapoptotic signaling events associated with life-threatening innate immune activation at the time of mammalian parturition.

  6. Surface Characterization of Virulent Treponema pallidum

    Science.gov (United States)

    Alderete, John F.; Baseman, Joel B.

    1980-01-01

    Characterization of the surface of Treponema pallidum was accomplished by [125I]lactoperoxidase-catalyzed iodination of intact organisms and sensitive radioimmunoprecipitation and gel electrophoresis technology. At least 11 outer membrane proteins with molecular weights ranging from 89,000 (89K) to 20K were identified, and all elicited high titers of antibody in experimentally infected rabbits. Proteins of 89.5K, 29.5K, and 25.5K previously implicated as ligands involved in attachment (J. B. Baseman and E. C. Hayes, J. Exp. Med. 151:573-586, 1980) were found to reside on the treponemal surface. Low levels of the 89.5K treponemal protein were released by high salt concentrations, whereas the remaining comigrating material was neither radioiodinated nor released with selective detergents. Other lower-molecular-weight (60K, 45K, and 30K) surface proteins were extracted with octyl glucoside detergent, suggesting their hydrophobic interaction with the external membrane. The molecular organization of surface proteins was studied by employing the cross-linker dithiobis(succinimidyl)-propionate, and data suggested the presence of a highly fluid envelope resulting in random collisions by the surface proteins. The biological function of the treponemal outer envelope proteins was evaluated using, as the indicator system, adherence of T. pallidum to monolayer cultures of eucaryotic cells. Trypsin treatment of motile, freshly harvested organisms decreased the extent of surface parasitism to normal rabbit testicular cells, reinforcing the idea of the proteinaceous nature and role of treponemal ligands for attachment. Other data supported functional and antigenic relatedness among the implicated ligands. Finally, brief periodate treatment of human epithelial (HEp-2) and normal rat testicular cells as well as casein-elicited rabbit peritoneal macrophages significantly reduced the extent of treponemal parasitism, suggesting a role of specific host membrane molecules as mediators of

  7. Synthesis, characterization and crystal structure determination of Mn (II) ion based 1D polymer constructed from 2, 2‧ bipyridyl and azide group, its thermal stability, magnetic properties and Hirshfeld surface analysis

    Science.gov (United States)

    Mudsainiyan, R. K.; Jassal, Amanpreet Kaur; Chawla, S. K.

    2015-05-01

    The 1-D polymeric complex (I) is having formula [Mn(2,2‧-BP).(N3)2]n, which has been crystallized in distilled water and characterized by elemental analyses, FT-IR spectrum, powder X-ray diffraction analyses and single-crystal diffraction analysis. This polymer possesses 1D helical chains or coils where Mn-azide-Mn forms the base of the coil which is alternatively garlanded by rigid bi-pyridine rings, where coordinates are in anti-fashion. The Mn (II) ions in the repeating units are linked by two end-on azide groups which extend through the two end-to-end azide ligands to the next unit forming a 1-D polymeric chain. The present study suggests that the use of this rigid and neutral building block leads to give better arrangement of the polymeric motif with [010] chains in 2-c uninodal net. During investigation of strong or weak intermolecular interactions, X-ray diffraction analysis and Hirshfeld surface analysis give rise to comparable results but in Hirshfeld surface analysis, two-third times more results of close contacts are obtained. The fingerprint plots demonstrate that these weak non-bonding interactions are important for stabilizing the crystal packing. Magnetic properties of the complex (I) were analyzed on the basis of an alternating ferro- and antiferromagnetic Heisenberg chain of Mn (II) ions. The J-exchange parameters found are J1=64.3 K (45.3 cm-1), and J2=-75.7 K (-53.3 cm-1). Magnetic properties are discussed in comparison with those of other similar molecular magnets of [Mn(L-L)(N3)2]n type.

  8. Zinc oxide Chemical Bath Deposition on Functionalized organic thin films: Formation of nanorods, nanorockets and nanoflowers

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Zhiwei; Walker, Amy V., E-mail: amy.walker@utdallas.edu

    2016-05-01

    Controlling the morphology of nanostructured materials is critical for their use in technological applications including in sensing, electronics and energy harvesting. In this paper we investigate the reaction pathways involved and their dependence on reactant concentrations in the formation of ZnO nanomaterials on –COOH terminated self-assembled monolayers using a simple chemical bath deposition process which employs zinc acetate, which acts as the Zn source, and ethylenediamine, which acts as both the O source and a complexing agent for Zn{sup 2+}. At a deposition temperature of 318 K (45 °C) our data shows that the concentration of Zn{sup 2+} as well as the deposition bath pH, which is controlled by the ethylenediamine concentration, is critical in determining the ZnO morphology. Above 0.01 M zinc acetate at low bath pH (~ 7.7–8.5), nanorods and nanorockets are observed to form. The nanorods exhibit a clear interface in the middle indicating that they are composed of two crystals. At lower zinc acetate concentrations over a wide pH range (~ 8.0–10.5) nanoflowers form. The nanorockets and nanoflowers grow via a modified La Mer mechanism in which there are multiple nucleation and crystallization steps. The initial nuclei are sphelurites (nanoflowers) or nanocrystallites (nanorockets). Since the reagent concentrations limit the reaction, for these initial precursor crystallites to increase in size, it is required dissolution and re-precipitation must occur. Thus at later times nanorockets or nanoflowers develop. - Highlights: • Nanorods, nanorockets and nanoflowers form depending on the reaction conditions. • Nanorods grow slowly suggesting low supersaturation conditions are needed. • Nanoflowers and nanorockets form via multiple nucleation and precipitation steps.

  9. Paleomagnetism of Eocene red-beds in the eastern part of the Qiangtang Terrane and its implications for uplift and southward crustal extrusion in the southeastern edge of the Tibetan Plateau

    Science.gov (United States)

    Tong, Yabo; Yang, Zhenyu; Mao, Changping; Pei, Junling; Pu, Zongwen; Xu, Yingchao

    2017-10-01

    A primary magnetic component with the acquisition time of 56.0-43.2 Ma was isolated between 580 °C and 685 °C from the Eocene Gonjo and Ranmugou Formations in the eastern part of the Qiangtang Terrane, Tibetan Plateau. The tilt corrected site-mean direction is Ds = 35.5 °, Is = 29.3 °, k = 45.9, α95 = 3.2 °. The site-mean inclination increased from 29.3° to 41.6° after multiple inclination shallowing corrections, giving a paleopole of 57.9°N/192.1°E, A95 = 2.9 °. Comparison of the Paleogene paleomagnetic data for the Qiangtang Terrane and Lhasa Terrane reveals that both terranes experienced latitudinal crustal shortening before 54-43 Ma, indicating the uplift of southern and central Tibet in the early Eocene. Subsequently, since 35.4 ± 2.4 Ma, the north of the Qiangtang Terrane experienced ∼1300 ± 410 km of crustal shortening, indicating the uplift of the northern Tibetan Plateau. The Lhasa Terrane and Qiangtang Terrane have not experienced further crustal shortening since the late Eocene, and the southeastern part of Tibet cannot have provided abundant crustal material to accommodate the significant crustal southward extrusion in the southeastern edge of the Tibetan Plateau. The crust of the Tengchong Terrane and Shan-Thai Block did not experience significant southward extrusion since the late Oligocene-Early Miocene. The Indochina Block was situated in the north of the Qiangtang Terrane before the Oligocene, and since the early Oligocene, the Indochina Block began to experience southward extrusion from the north of the Qiangtang Terrane, which absorbed part of the crustal shortening in the north of the Qiangtang Terrane.

  10. A human vitamin D receptor mutation causes rickets and impaired Th1/Th17 responses.

    Science.gov (United States)

    van der Eerden, Bram C J; van der Heyden, Josine C; van Hamburg, Jan Piet; Schreuders-Koedam, Marijke; Asmawidjaja, Patrick S; de Muinck Keizer-Schrama, Sabine M; Boot, Annemieke M; Lubberts, Erik; Drop, Stenvert L S; van Leeuwen, Johannes P T M

    2014-12-01

    We present a brother and sister with severe rickets, alopecia and highly elevated serum levels of 1,25-dihydroxyvitamin D (1,25-(OH)2D3). Genomic sequencing showed a homozygous point mutation (A133G) in the vitamin D receptor gene, leading to an amino acid change in the DNA binding domain (K45E), which was described previously. Hereditary vitamin D resistant rickets (HVDRR) was diagnosed. Functional studies in skin biopsy fibroblasts confirmed this. 1,25-(OH)2D3 reduced T helper (Th) cell population-specific cytokine expression of interferon γ (Th1), interleukins IL-17A (Th17) and IL-22 (Th17/Th22) in peripheral blood mononuclear cells (PBMCs) from the patient's parents, whereas IL-4 (Th2) levels were higher, reflecting an immunosuppressive condition. None of these factors were regulated by 1,25-(OH)2D3 in PBMCs from the boy. At present, both patients (boy is 23 years of age, girl is 7) have not experienced any major immune-related disorders. Although both children developed alopecia, the girl did so earlier than the boy. The boy showed complete recovery from the rickets at the age of 17 and does not require any vitamin D supplementations to date. In conclusion, we characterized two siblings with HVDRR, due to a mutation in the DNA binding domain of VDR. Despite a defective T cell response to vitamin D, no signs of any inflammatory-related abnormalities were seen, thus questioning an essential role of vitamin D in the immune system. Despite the fact that currently medicine is not required, close monitoring in the future of these patients is warranted for potential recurrence of vitamin D dependence and diagnosis of (chronic) inflammatory-related diseases.

  11. Assessment of natural radioactivity levels in rocks and their relationships with the geological structure of Johor state, Malaysia.

    Science.gov (United States)

    Alnour, I A; Wagiran, H; Ibrahim, N; Hamzah, S; Elias, M S; Laili, Z; Omar, M

    2014-01-01

    The distribution of natural radionuclides ((238)U, (232)Th and (40)K) and their radiological hazard effect in rocks collected from the state of Johor, Malaysia were determined by gamma spectroscopy using a high-purity germanium detector. The highest values of (238)U, (232)Th and (40)K activity concentrations (67±6, 85±7 and 722±18 Bg kg(-1), respectively) were observed in the granite rock. The lowest concentrations of (238)U and (232)Th (2±0.1 Bq kg(-1) for (238)U and 2±0.1 Bq kg(-1) for (232)Th) were observed in gabbro rock. The lowest concentration of (40)K (45±2 Bq kg(-1)) was detected in sandstone. The radium equivalent activity concentrations for all rock samples investigated were lower than the internationally accepted value of 370 Bq kg(-1). The highest value of radium equivalent in the present study (239±17 Bq kg(-1)) was recorded in the area of granite belonging to an acid intrusive rock geological structure. The absorbed dose rate was found to range from 4 to 112 nGy h(-1). The effective dose ranged from 5 to 138 μSv h(-1). The internal and external hazard index values were given in results lower than unity. The purpose of this study is to provide information related to radioactivity background levels and the effects of radiation on residents in the study area under investigation. Moreover, the relationships between the radioactivity levels in the rocks within the geological structure of the studied area are discussed.

  12. Visualization of Uptake of Mineral Elements and the Dynamics of Photosynthates in Arabidopsis by a Newly Developed Real-Time Radioisotope Imaging System (RRIS).

    Science.gov (United States)

    Sugita, Ryohei; Kobayashi, Natsuko I; Hirose, Atsushi; Saito, Takayuki; Iwata, Ren; Tanoi, Keitaro; Nakanishi, Tomoko M

    2016-04-01

    Minerals and photosynthates are essential for many plant processes, but their imaging in live plants is difficult. We have developed a method for their live imaging in Arabidopsis using a real-time radioisotope imaging system. When each radioisotope,(22)Na,(28)Mg,(32)P-phosphate,(35)S-sulfate,(42)K,(45)Ca,(54)Mn and(137)Cs, was employed as an ion tracer, ion movement from root to shoot over 24 h was clearly observed. The movements of(22)Na,(42)K,(32)P,(35)S and(137)Cs were fast so that they spread to the tip of stems. In contrast, high accumulation of(28)Mg,(45)Ca and(54)Mn was found in the basal part of the main stem. Based on this time-course analysis, the velocity of ion movement in the main stem was calculated, and found to be fastest for S and K among the ions we tested in this study. Furthermore, application of a heat-girdling treatment allowed determination of individual ion movement via xylem flow alone, excluding phloem flow, within the main stem of 43-day-old Arabidopsis inflorescences. We also successfully developed a new system for visualizing photosynthates using labeled carbon dioxide,(14)CO2 Using this system, the switching of source/sink organs and phloem flow direction could be monitored in parts of whole shoots and over time. In roots,(14)C photosynthates accumulated intensively in the growing root tip area, 200-800 µm behind the meristem. These results show that this real-time radioisotope imaging system allows visualization of many nuclides over a long time-course and thus constitutes a powerful tool for the analysis of various physiological phenomena.

  13. Methodology of Ecological Evaluation and Regulation of Soil Degradation on the Basis of Equation for the Soil State Function

    Science.gov (United States)

    Evdokimova, Mariya; Vladimir, Gendugov; Gennady, Glazunov

    2016-04-01

    Procedures of determining threshold concentrations of stressors on ecosystem components (for example, maximum permissible concentration (MPC)) include experimental study of responses to impacts. Full curves of the response of living organisms and populations have, as a rule, graphics in the form of a deformed bell and, in the absence of theoretical models, are interpreted either, using experimental or statistical models, which reduces the information content of experimental data. More preferably to use a theoretical model, free of the drawbacks of empirical models: q = C/zb∗exp(-K/z), (1) where q - is a measurable response of living organisms on exposure to the stressor, the concentration of which is equal to z,C - the constant of integration that makes sense of coefficient, which is scaling the value q , b-the coefficient of velocity of growth with the increase of z,K - the coefficient of velocity decrease q with increasing z. The coefficients C,b,Kof equation (1) find by fitting the model (1) to the experimental data by methods of nonlinear regression using an available software package. The algorithm involves the introduction into the program of approximate values of the coefficients. These approximate values calculated by analytical formulas, obtained from (1): lnC = (c3∗a1∗b2-c3∗b1∗a2-c1∗a3∗b2-c2∗a1∗b3+c2∗b1∗a3+c1∗a2∗b3)/(b2∗a1-b1∗a2-a3∗b2-b3∗a1+b1∗a3+a2∗b3), (2) b = (c2 ∗ a1 - c1 ∗ a2+(a2-a1)*lnC)/(b2∗a1-b1∗a2), (3) K = (c1 - b ∗ b1-lnC)/a1, (4) where ai = -1/zi,bi = -lnzi,ci =lnqi,qi-the current value of the response of living organisms to the impact of the stressor at a concentration of zi . For the calculations three pairs of representative values are chosen from the experimental data, and substituted into equation (2) - (6). Using the obtained constants C,b,K, values of qi are calculated, using equation (1), for all zifrom the table and the average relative deviation of calculated values qifrom measured , delta is found. The definition of constants is repeated with different pairs of representative values qiand zi, achieving the minimum value of the average relative deviation delta. For the evaluation of the true values adopt those b and K, which correspond to the smallest value of delta.They are used to calculate the abscissa of the maximum point of the dose-response curve, based on the theoretical formula, obtained from the analysis of the first derivative dq/dz: zmax = K/b(5) and the abscissa of the inflection point of the graph (to the right of the maximum) according to the theoretical formula, obtained from the analysis of the second derivative d2q/dz2: zinf=(K*b+K+(b+1)1/2)/(b2 + b) (6) Acknowledgments: This study was supported by the Russian Science Foundation, project no. 143800023.

  14. Hydromechanical Simulations of Surface Uplift due to CO2 Injection at In Salah (Invited)

    Science.gov (United States)

    Morris, J. P.; Hao, Y.; Foxall, W.; McNab, W. W.

    2009-12-01

    We present recent simulations of the hydromechanical response of the reservoir and overburden associated with CO2 injection at In Salah. Using the best available field data for the reservoir and fault network properties, we are able to demonstrate excellent agreement between simulation and observation. These results are providing new insight into the fate of the CO2 about one of the injectors where intriguing morphology was observed in surface uplift. Additionally, this work is helping to better establish the advantages and limitations of interpreting surface displacements to guide our understanding of fluid fate. The In Salah Project (a joint venture of BP, StatoilHydro and Sonatrach) includes a CO2 sequestration effort that has successfully injected millions of tons of CO2 into a deep saline formation close to a producing gas field in Algeria. We have been funded by the Joint Industry Project (A consortium consisting of BP, StatoilHydro and Sonatrach, hereafter the JIP) and the U.S. Department of Energy to investigate the role of injection induced mechanical deformation and geochemical alteration at the In Salah CO2 storage project. Here we focus upon the hydromechanical portion of the study. We have performed detailed simulations of the hydromechanical response in the vicinity of the KB-502 CO2 injector specifically because the morphology of the observed surface deformation differed from that above the other injectors at the field. First we performed a geomechanical analysis to predict which faults are flow conduits and which are flow barriers. NUFT simulations were performed based upon this information using permeability fields for the reservoir provided by the JIP. These results indicate that the presence of faults in the vicinity of the KB-502 injector may be responsible for the early breakthrough of CO2 observed at a nearby well, KB-5. We have simulated the mm-scale uplift of the overburden and compared the results with observed deformation using InSAR data

  15. Heat capacity of paramagnetic nickelocene: Comparison with diamagnetic ferrocene

    Science.gov (United States)

    Sorai, Michio; Kaneko, Yuki; Hashiguchi, Takao

    2014-05-01

    Nickelocene [bis(η5-cyclopentadienyl)nickel: Ni(C5H5)2, electron spin S=1, the ground state configuration 3A2g] is paramagnetic and belongs to a typical molecule-based magnet. Heat capacities of nickelocene have been measured at temperatures in the 3-320 K range by adiabatic calorimetry. By comparing with those of diamagnetic ferrocene crystal, a small heat capacity peak centered at around 15 K and a sluggish hump centered at around 135 K were successfully separated. The low-temperature peak at 15 K caused by the spin is well reproduced by the Schottky anomaly due to the uniaxial zero-field splitting of the spin S=1 with the uniaxial zero-field splitting parameter D/k=45 K (k: the Boltzmann constant). The magnetic entropy 9.7 J K-1mol-1 is substantially the same as the contribution from the spin-manifold R ln 3=9.13 J K-1mol-1 (R: the gas constant). The sluggish hump centered at around 135 K arises from rotational disordering of the cyclopentadienyl rings of nickelocene molecule. The enthalpy and entropy gains due to this anomaly are 890 J mol-1 and 6.9 J K-1mol-1, respectively. As the hump spreads over a wide temperature region, separation of the hump from the observed heat capacity curve involves a little bit ambiguity. Therefore, these values should be regarded as being reasonable but tentative. The present entropy gain is comparable with 5.5 J K-1mol-1 for the sharp phase transition at 163.9 K of ferrocene crystal. This fact implies that although the disordering of the rings likewise takes place in both nickelocene and ferrocene, it proceeds gradually in nickelocene and by way of a cooperative phase transition in ferrocene. A reason for this originates in loose molecular packing in nickelocene crystal. Molar heat capacity and the standard molar entropy of nickelocene are larger than those of ferrocene beyond the mass effect over the whole temperature region investigated. This fact provides with definite evidences for the loose molecular packing in nickelocene

  16. Dual Vulnerability of Tau to Calpains and Caspase-3 Proteolysis Under Neurotoxic and Neurodegenerative Conditions

    Directory of Open Access Journals (Sweden)

    Ming Cheng Liu

    2010-11-01

    Full Text Available Axonally specific microtubule-associated protein tau is an important component of neurofibrillary tangles found in AD (Alzheimer's disease and other tauopathy diseases such as CTE (chronic traumatic encephalopathy. Such tau aggregate is found to be hyperphosphorylated and often proteolytically fragmented. Similarly, tau is degraded following TBI (traumatic brain injury. In the present study, we examined the dual vulnerability of tau to calpain and caspase-3 under neurotoxic and neurodegenerative conditions. We first identified three novel calpain cleavage sites in rat tau (four-repeat isoform as Ser130 ↓ Lys131, Gly157 ↓ Ala158 and Arg380 ↓ Glu381. Fragment-specific antibodies to target the major calpain-mediated TauBDP-35K (35 kDa tau-breakdown product and the caspase-mediated TauBDP-45K respectively were developed. In rat cerebrocortical cultures treated with excitotoxin [NMDA (N-methyl-D-aspartate], tau is significantly degraded into multiple fragments, including a dominant signal of calpain-mediated TauBDP-35K with minimal caspase-mediated TauBDP-45K. Following apoptosis-inducing EDTA treatment, tau was truncated only to TauBDP-48K/45K-exclusively by caspase. Cultures treated with another apoptosis inducer STS (staurosporine, dual fragmentation by calpain (TauBDP-35K and caspase-3 (TauBDP-45K was observed. Tau was also fragmented in injured rat cortex following TBI in vivo to BDPs of 45-42 kDa (minor, 35 kDa and 15 kDa, followed by TauBDP-25K. Calpain-mediated TauBDP-35K-specific antibody confirmed robust signals in the injured cortex, while caspase-mediated TauBDP-45K-specific antibody only detected faint signals. Furthermore, intravenous administration of a calpain-specific inhibitor SNJ-1945 strongly suppressed the TauBDP-35K formation. Taken together, these results suggest that tau protein is dually vulnerable to calpain and caspase-3 proteolysis under different neurotoxic and injury conditions.

  17. Comprehensive enzymatic analysis of the cellulolytic system in digestive fluid of the Sea Hare Aplysia kurodai. Efficient glucose release from sea lettuce by synergistic action of 45 kDa endoglucanase and 210 kDa ß-glucosidase.

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    Akihiko Tsuji

    Full Text Available Although many endo-ß-1,4-glucanases have been isolated in invertebrates, their cellulolytic systems are not fully understood. In particular, gastropod feeding on seaweed is considered an excellent model system for production of bioethanol and renewable bioenergy from third-generation feedstocks (microalgae and seaweeds. In this study, enzymes involved in the conversion of cellulose and other polysaccharides to glucose in digestive fluids of the sea hare (Aplysia kurodai were screened and characterized to determine how the sea hare obtains glucose from sea lettuce (Ulva pertusa. Four endo-ß-1,4-glucanases (21K, 45K, 65K, and 95K cellulase and 2 ß-glucosidases (110K and 210K were purified to a homogeneous state, and the synergistic action of these enzymes during cellulose digestion was analyzed. All cellulases exhibited cellulase and lichenase activities and showed distinct cleavage specificities against cellooligosaccharides and filter paper. Filter paper was digested to cellobiose, cellotriose, and cellotetraose by 21K cellulase, whereas 45K and 65K enzymes hydrolyzed the filter paper to cellobiose and glucose. 210K ß-glucosidase showed unique substrate specificity against synthetic and natural substrates, and 4-methylumbelliferyl (4MU-ß-glucoside, 4MU-ß-galactoside, cello-oligosaccharides, laminarin, and lichenan were suitable substrates. Furthermore, 210K ß-glucosidase possesses lactase activity. Although ß-glucosidase and cellulase are necessary for efficient hydrolysis of carboxymethylcellulose to glucose, laminarin is hydrolyzed to glucose only by 210K ß-glucosidase. Kinetic analysis of the inhibition of 210K ß-glucosidase by D-glucono-1,5-lactone suggested the presence of 2 active sites similar to those of mammalian lactase-phlorizin hydrolase. Saccharification of sea lettuce was considerably stimulated by the synergistic action of 45K cellulase and 210K ß-glucosidase. Our results indicate that 45K cellulase and 210K

  18. Synthesis, characterization and crystal structure determination of Mn (II) ion based 1D polymer constructed from 2, 2′ bipyridyl and azide group, its thermal stability, magnetic properties and Hirshfeld surface analysis

    Energy Technology Data Exchange (ETDEWEB)

    Mudsainiyan, R.K., E-mail: mudsainiyanrk@gmail.com; Jassal, Amanpreet Kaur; Chawla, S.K., E-mail: sukhvinder.k.chawla@gmail.com

    2015-05-15

    The 1-D polymeric complex (I) is having formula [Mn(2,2′-BP).(N{sub 3}){sub 2}]{sub n}, which has been crystallized in distilled water and characterized by elemental analyses, FT-IR spectrum, powder X-ray diffraction analyses and single-crystal diffraction analysis. This polymer possesses 1D helical chains or coils where Mn–azide–Mn forms the base of the coil which is alternatively garlanded by rigid bi-pyridine rings, where coordinates are in anti-fashion. The Mn (II) ions in the repeating units are linked by two end-on azide groups which extend through the two end-to-end azide ligands to the next unit forming a 1-D polymeric chain. The present study suggests that the use of this rigid and neutral building block leads to give better arrangement of the polymeric motif with [010] chains in 2-c uninodal net. During investigation of strong or weak intermolecular interactions, X-ray diffraction analysis and Hirshfeld surface analysis give rise to comparable results but in Hirshfeld surface analysis, two-third times more results of close contacts are obtained. The fingerprint plots demonstrate that these weak non-bonding interactions are important for stabilizing the crystal packing. Magnetic properties of the complex (I) were analyzed on the basis of an alternating ferro- and antiferromagnetic Heisenberg chain of Mn (II) ions. The J-exchange parameters found are J{sub 1}=64.3 K (45.3 cm{sup −1}), and J{sub 2}=−75.7 K (−53.3 cm{sup −1}). Magnetic properties are discussed in comparison with those of other similar molecular magnets of [Mn(L–L)(N{sub 3}){sub 2}]{sub n} type. - - Highlights: • Synthesized 1-D polymeric complex of Mn (II) ions with 2, 2′ bipyridyl and azide group. • X-ray data of complex (I) is in a good agreement with TGA and other spectroscopic techniques. • DFT calculations were done and compared with the parameter of experimental and theoretical data. • Intermolecular interactions calculated by Hirshfeld surface analysis

  19. Bacterial effector binding to ribosomal protein s3 subverts NF-kappaB function.

    Directory of Open Access Journals (Sweden)

    Xiaofei Gao

    2009-12-01

    as an AP-1-dependent reporter. We identified a region of NleH1 (N40-K45 that is at least partially responsible for the inhibitory activity of NleH1 toward RPS3. Deleting nleH1 from E. coli O157:H7 produced a hypervirulent phenotype in a gnotobiotic piglet model of Shiga toxin-producing E. coli infection. We suggest that NleH may disrupt host innate immune responses by binding to a cofactor of host transcriptional complexes.

  20. Effect of high soil copper concentration on mycorrhizal grapevines

    Science.gov (United States)

    Nogales, Amaia; Santos, Erika S.; Viegas, Wanda; Aran, Diego; Pereira, Sofia H.; Vidigal, Patricia; Lopes, Carlos M.; Abreu, M. Manuela

    2017-04-01

    Repeated application of Copper (Cu) based fungicides in vineyards since the end of the 19th century has led to a significant increase in the concentration of this chemical element in many viticultural soils. Although Cu is an essential micronutrient for most organisms, it can be toxic for the development and survival of plants and soil (micro)organisms at high concentrations and eventually lead to yield loses in viticulture, as it negatively affects key physiological and biogeochemical processes. However, some soil microorganisms, including arbuscular mycorrhizal fungi (AMF), have developed adaptive mechanisms for persistence in environments with supra-optimal levels of essential elements or in the presence of harmful ones, as well as for increasing plant tolerance to such abiotic stress conditions. The objective of this work was to evaluate the effect of a high total soil concentration of Cu on microbial soil activity as well as on the development of mycorrhizal and non-mycorrhizal grapevines. A microcosm assay was set up under greenhouse and controlled conditions. Touriga Nacional grapevine variety plants grafted onto 1103P rootstocks were inoculated either with the AMF Rhizophagus irregularis or Funneliformis mosseae, or were left as non-inoculated controls. After three months, they were transplanted to containers filled with 4 kg of a sandy soil (pH: 7.0; electrical conductivity: 0.08 mS/cm; [organic C]: 5.6 g/kg; [N-NO3]: 1.1 mg/kg; [N-NH4]: 2.5 mg/kg; [extractable K]: 45.1 mg/kg; [extractable P]: 52.3 mg/kg), collected near to a vineyard in Pegões (Portugal). Two treatments were carried out: with and without Cu application. The soil with high Cu concentration was prepared by adding 300 mg Cu/kg (in the form of an aqueous solution of CuSO4·5H2O) followed by an incubation during four weeks in plastic bags at room temperature in dark. Physico-chemical soil characteristics (pH, electrical conductivity and nutrients concentration in available fraction), soil

  1. Bacterial effector binding to ribosomal protein s3 subverts NF-kappaB function.

    Directory of Open Access Journals (Sweden)

    Xiaofei Gao

    2009-12-01

    as an AP-1-dependent reporter. We identified a region of NleH1 (N40-K45 that is at least partially responsible for the inhibitory activity of NleH1 toward RPS3. Deleting nleH1 from E. coli O157:H7 produced a hypervirulent phenotype in a gnotobiotic piglet model of Shiga toxin-producing E. coli infection. We suggest that NleH may disrupt host innate immune responses by binding to a cofactor of host transcriptional complexes.

  2. Epidemiologic Features and Prognosis of Patients Who Was Diagnosed Having Type 2 Diabetes Mellitus and Applied to a Community Health Center

    Directory of Open Access Journals (Sweden)

    Sevilay Hindistan

    2009-08-01

    Full Text Available AIM: The study was designed to investigate the prognosis and the epidemiologic features of people who was diagnosed having type 2 DM and Applied to the Community Health Center. METHOD: This study which was carried out at Catak Health Branch of Trabzon Erdogdu Health Center in April, May, and June in 2008 is a descriptive and cross-sectional field study. All of the patients with Type 2 DM diagnosis who came to the Health Center for routine control (55 people were involved in the study. The aim of the study was explained to each participant in the study and their oral consent was taken. As the data collection instrument, diabetes surveillance form which includes of socio-demographic features, risk factors, blood sugar diagnosis criteria, body mass index, chronic complications and foot examination was used. Diabetes surveillance form was shaped by modifying the diabetes diagnosis form which was structured by Erdogan and Nahcivan (1999. The data obtained were evaluated through number and percentage distribution, mean, chi square test, correlation and variant analyses techniques. RESULTS: Age mean of the patients was 59.4±10.7 and 85.5% of them was female. Average diagnosis age was 53.2±12.5, average diabetes year was 6,6±6,6. 45.5% of the patients were obese, 12.7% of them morbid obese and diabetes history in the family was about 54.5%. It was found that 56.3% patients had blood glucose level was 140 mg/dl and 54.5% of the patients’ HbA1c level were higher than 7%. When lipid parameter levels of the patients were examined, it was seen that HDL cholesterol level was (E<35,K<45 for 52.7% of the patients and that was lower than the target treatment level; LDL cholesterol level was 100mg/dl for 76.4%; and total cholesterol level was higher than 200 mg/dl 49.1% of the patients. A statistically significant positive correlation was found between diabetes year of the patients and HbA1c level (r=0.291, p=0.031. 8.6 had % neuropathy, 1.7% of them had

  3. Private sports centers in Ankara task levels of job satisfaction of staff engaged in evaluationAnkara’daki özel spor merkezlerinde çalışan personelin iş doyum düzeylerinin değerlendirilmesi

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    Yalçın Uyar

    2016-08-01

    Full Text Available The aim of this study is to assess job satisfaction levels of the personnel that is working for private sports centers in Ankara. Approximately 700 personnel working for 45 private sports centers in Ankara having at least 400 members constitute universe of the research which is held according to descriptive research model. Also, 87 female (%50,6 and 85 male (%49,4 making total of 172 personnel composed work group of the research. As data collection tool, Balcı (1985 is used to define job satisfaction scale developed by the private sports center staff. Analysis of the data standard deviation, arithmetic means, that we look at the factor analysis and item-total correlations, analysis of variance to compare the differences between groups and t-test was used. In the safety study for overall Cronbach's Alpha coefficient was calculated to be .945. As a result of the work of private sports center staff it has been shown to provide great satisfaction rate.   Özet Bu araştırmanın amacı,Ankara’daki özel spor merkezlerinde çalışan personelin iş doyum düzeylerinin değerlendirilmesidir.Betimsel ilişkisel tarama modeline uygun olarak düzenlenen araştırmanın evrenini,Ankara’da,en az 400 üyesi bulunan yaklaşık 45 özel spor merkezinde görev yapan yaklaşık 700 çalışan oluşturmaktadır.Çalışma grubunu ise,anılan  evren içerisinde tesadüfi yöntem ile seçilmiş Ankara ilindeki özel spor merkezlerinin değişik kademelerinde görev yapan 87 kadın(% 50,6,85 erkek(%49,4 olmak üzere toplam 172 kişi oluşturmaktadır.Araştırmada veri toplama aracı olarak, Balcı(1985tarafından geliştirilen iş doyum ölçeği özel spor merkezi çalışanlarına uyarlanarak  kullanılmıştır. Verilerin analizinde standart sapma, aritmetik ortalamalar, faktör analizi ve madde–toplam test korelasyonuna bakılmış, grup ortalamaları arasındaki farklılıkları karşılaştırmak için varyans analizi ve t-testi kullan

  4. 长期施肥红壤钾素在有机无机复合体中的分布%Distribution of potassium in the organo-mineral complexes of red soils under long-term fertilization

    Institute of Scientific and Technical Information of China (English)

    岳龙凯; 蔡泽江; 徐明岗; 王伯仁; 黄庆海; 李冬初; 柳开楼; 李建军; 张会民

    2015-01-01

    生长的主要钾源,其中<2 μm粒级复合体中交换性钾、非交换性钾含量与产量之间均存在显著正相关关系( P<0.05 ). [结论] <2 μm粒级复合体是土壤钾素的主要贮存库. 在长期不施钾肥条件下土壤<2 μm粒级复合体中交换性钾和非交换性钾含量降低,施钾有利于该粒级复合体中交换性钾和非交换性钾积累. 旱地红壤<2、2 10及10 50 μm粒级复合体钾素含量高,且与产量之间存在显著的正相关关系,是植物钾素主要供源.%[Objectives] Soil organo-mineral complexes contain most of soil nutrients that have become a most important part of soil nutrients .Change of soil potassium contents affects the contents of potassium in different sizes of organo-mineral complex .So long-term experiments of red soil started from 1990 ( Qiyang ) and 1986 ( Jinxian ) were carried out to investigate distribution of exchangeable , non-exchangeable and total potassium ( K) in different sizes of organo-mineral complex .[Methods] The soil samples of 0-20 cm depth were collected from the four treatments in the long term experiment at both sites:No fertilization ( CK) , mixed application of chemical nitrogen and phosphorus fertilizer ( NP ) , mixed application of NP combined with chemical potassium fertilizer ( NPK ) , mixed application of NPK combined with manure ( NPKM ) treatments. Stokes law was used to calculate sedimentation rates of different sizes of organo-mineral complex which were separated using siphon .The soil exchangeable, non-exchangeable and total potassium were determined .[Results] In the two sites, 96.4%-98.9%of the exchangeable K , 87.9%-96.7%of the non-exchangeable K and 95.1%-96.1%of the total K are in 0-50 μm sizes of organo-mineral complex .Especially , 76.3%-92.3% of the exchangeable K , 45.8%-73.7%of the non-exchangeable K and 49.4%-70.6% of the total K at both sites are mainly in <2 μm size complex.Comparing with CK, the contents of non-exchangeable K under NP are decreased by 5.4 to 8

  5. SUSTAINABILITY EFFECTS OF Crotalaria juncea L. AND Crotalaria spectabilis ROTH ON SOIL FERTILITY AND SOIL CONSERVATION

    Science.gov (United States)

    László, Márton, ,, Dr.

    2010-05-01

    ROTH) before flowering amounted to 368 kg N, 252 kg Ca, 96 kg K, 45 kg Mg, 30 kg P and 27 kg S ha-1. The content of Al and Fe total 2 - 3, while that of Ba, Zn, B, Cu, Na, Mn and Sr 180 - 650 g ha-1. The Co, Cd, As, Pb, Ni, Se, Cr and Mo concentration did not reach here the value of 10 g ha-1. By this means this green manures should have a vary important role in the design of rotations for sustainable agriculture. Not only do they help to retain and accumulate nitrogen and other nutrients, thus reducing leaching losses, they also maintain ground cover, protected the soil from erosion, and can make a contribution to pest and weed control. Key words: Sustainable agriculture, soil fertility, soil conservation, green manure, Crotalaria juncea L., Crotalaria spectabilis ROTH. INTRODUCTION Sustainable agriculture is defined as the successful management of resources for agriculture to satisfy changing human needs while maintaining or enhancing the quality of the environment and conserving natural resources. A sustained agricultural production can be achieved by a proper use of soil resources, which includes the maintance or the enhancement of soil fertility (Christian and Kurt 1996). The term soil fertility is cast here to encompass not only essential plant nutrients but also aspects of soil structure, including water holding capacity, soil organic matter content and biological activity that influence both the efficiency of use and sustainability of the resources. All these attributes are interrelated and contribute together to the soil potential productivity or fertility (Kádár 1992, Németh 1996). From that perspective, soil fertility can be assessed as a capital stock, which will produce interests when properly used , and yet will be eroded by a consumptive use. It is necessary to make here a clear distinction between actions aiming at the regeneration of the soil capital, i.e., "recapitalization of soil fertility" and actions, such as maintance of enhanced soil