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Sample records for unit e1 zim-sci

  1. 26 CFR 31.3121(e)-1 - State, United States, and citizen.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false State, United States, and citizen. 31.3121(e)-1... § 31.3121(e)-1 State, United States, and citizen. (a) When used in the regulations in this subpart, the... is used in a geographical sense. The term “citizen of the United States” includes a citizen of the...

  2. 26 CFR 1.927(e)-1 - Special sourcing rule.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 10 2010-04-01 2010-04-01 false Special sourcing rule. 1.927(e)-1 Section 1.927... (CONTINUED) INCOME TAXES Earned Income of Citizens of United States § 1.927(e)-1 Special sourcing rule. (a... under section 925 with respect to such transaction applied to such transaction. This special sourcing...

  3. The Implementation of E1 Clock Recovery

    Directory of Open Access Journals (Sweden)

    Wang Ziyu

    2016-01-01

    Full Text Available Clock transform and recovery is of significant importance in microwave TDM service, and it is always extracted from the E1 line data stream in most cases. However, intrinsically uncertain delay and jitter caused by packet transmission of E1 data information, may lead to the indexes of the data recovery clock exceed the clock performance template. Through analysis of the E1 clock indexes and measuring methods, this paper proposes a new clock recovery method. The method employs two buffers, the first RAM is used as a buffer to deduct excess information, and the second FIFO is used as a buffer to recovery the clock and data. The first buffer has a feedback from the second one, and is able to actively respond to changes in the data link and requests from the second one. The test results validate the effectiveness of the method, and the corresponding scheme is also valuable for the other communication systems.

  4. E1 strength in N = 82 nuclei

    Energy Technology Data Exchange (ETDEWEB)

    Hartmann, T.; Volz, S.; Babilon, M.; Mohr, P.; Vogt, K.; Zilges, A

    2003-05-19

    Recently the importance of small contributions of electric dipole strength near the particle threshold to the production rates of atomic nuclei has become evident. Prior estimates concentrated on the Giant Dipole Resonance (GDR) which dominates photoabsorption in all nuclei. Extrapolations to smaller excitation energies were assumed to be sufficiently reliable. However, new measurements reveal that collective E1 strength can be found in the threshold region.

  5. E1M1 and E1E2 transition probabilities in one-electron ions

    Science.gov (United States)

    Labzowsky, L. N.; Shonin, A. V.

    2004-12-01

    The quantum electrodynamical (QED) theory of the two-photon transitions in hydrogenlike ions is presented. The emission probability for 2s→2γ(E1)+1s transitions is calculated and compared to the results of the previous calculations. The emission probabilities 2p→γ(E1)+γ(E2)+1s and 2p→γ(E1)+γ(M1)+1s are also calculated for the nuclear charge Z values 1⩽Z⩽100. This is the first calculation of the two latter probabilities. The results are given in two different gauges.

  6. E1M1 and E1E2 transition probabilities in one-electron ions

    Energy Technology Data Exchange (ETDEWEB)

    Labzowsky, L.N. [Institute of Physics, St. Petersburg State University, Uljanovskaya 1, Petrodvorets, 198904 St. Petersburg (Russian Federation) and Petersburg Nuclear Physics Institute, Gatchina, 188350 St. Petersburg (Russian Federation)]. E-mail: leonti@landau.phys.spbu.ru; Shonin, A.V. [Institute of Physics, St. Petersburg State University, Uljanovskaya 1, Petrodvorets, 198904 St. Petersburg (Russian Federation)

    2004-12-06

    The quantum electrodynamical (QED) theory of the two-photon transitions in hydrogenlike ions is presented. The emission probability for 2s1/2->2{gamma}(E1)+1s1/2 transitions is calculated and compared to the results of the previous calculations. The emission probabilities 2p1/2->{gamma}(E1)+{gamma}(E2)+1s1/2 and 2p1/2->{gamma}(E1)+{gamma}(M1)+1s1/2 are also calculated for the nuclear charge Z values 1=

  7. Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication

    Directory of Open Access Journals (Sweden)

    Natascha Krömmelbein

    2016-02-01

    Full Text Available The human cytomegalovirus (HCMV replicates to high titers in primary human fibroblast cell cultures. A variety of primary human cells and some tumor-derived cell lines do also support permissive HCMV replication, yet at low levels. Cell lines established by transfection of the transforming functions of adenoviruses have been notoriously resistant to HCMV replication and progeny production. Here, we provide first-time evidence that a permanent cell line immortalized by adenovirus type 5 E1A and E1B (CAP is supporting the full HCMV replication cycle and is releasing infectious progeny. The CAP cell line had previously been established from amniotic fluid cells which were likely derived from membranes of the developing fetus. These cells can be grown under serum-free conditions. HCMV efficiently penetrated CAP cells, expressed its immediate-early proteins and dispersed restrictive PML-bodies. Viral DNA replication was initiated and viral progeny became detectable by electron microscopy in CAP cells. Furthermore, infectious virus was released from CAP cells, yet to lower levels compared to fibroblasts. Subviral dense bodies were also secreted from CAP cells. The results show that E1A/E1B expression in transformed cells is not generally repressive to HCMV replication and that CAP cells may be a good substrate for dense body based vaccine production.

  8. (E-1-Benzylidene-2,2-diphenylhydrazine

    Directory of Open Access Journals (Sweden)

    Angel Mendoza

    2012-02-01

    Full Text Available The asymmetric unit of the title compound, C19H16N2, contains two independent molecules, both of which show an E configuration with respect to the C=N bond. The dihedral angles between the phenyl rings bonded to the hydrazine group are 81.00 (10 and 88.34 (8° in the two molecules. Intermolecular C—H...π interactions are observed in the crystal structure.

  9. (E-1-(3-Ethoxy-2-hydroxybenzylidenethiosemicarbazide

    Directory of Open Access Journals (Sweden)

    Amir Adabi Ardakani

    2012-02-01

    Full Text Available The title compound, C10H13N3O2S, crystallizes with two independent molecules (A and B in the asymmetric unit. In the crystal, the A and B molecules are linked via pairs of N—H...O and O—H...S hydrogen bonds, forming dimers with R22(14 and R22(6 ring motifs. These dimers are linked via a pair of N—H...S hydrogen bonds with an R22(8 ring motif, forming chains propagating along the c-axis direction. The crystal was refined as an inversion twin with a final BASF ratio of 0.54 (11:0.46 (11.

  10. Main: 1E1E [RPSD[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1E トウモロコシ Corn Zea mays L. Beta-Glucosidase, Chloroplast Precursor Name=Glu1; Zea...TDDAYASQEVNGPDGKPIGPPMGNPWIYMYPEGLKDLLMIMKNKYGNPPIYITENGIGDVDTKETPLPMEAALNDYKRLDYIQRHIATLKESIDLGSNVQGYFAWSLLDNFEWFAGFTERYGIVYVDRNNNCTRYMKESAKWLKEFNTAKKPSKKILTPA corn_1E1E.jpg ...

  11. Main: 1E1F [RPSD[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1F トウモロコシ Corn Zea mays L. Beta-Glucosidase, Chloroplast Precursor Name=Glu1; Zea...QRHIATLKESIDLGSNVQGYFAWSLLDNFEWFAGFTERYGIVYVDRNNNCTRYMKESAKWLKEFNTAKKPSKKILTPA corn_1E1F.jpg ...

  12. CYP2E1 Metabolism of Styrene Involves Allostery

    OpenAIRE

    2012-01-01

    We are the first to report allosterism during styrene oxidation by recombinant CYP2E1 and human liver microsomes. At low styrene concentrations, oxidation is inefficient because of weak binding to CYP2E1 (Ks = 830 μM). A second styrene molecule then binds CYP2E1 with higher affinity (Kss = 110 μM) and significantly improves oxidation to achieve a kcat of 6.3 nmol · min−1 · nmol CYP2E1−1. The transition between these metabolic cycles coincides with reported styrene concentrations in blood from...

  13. CYP2E1 autoantibodies in liver diseases

    Science.gov (United States)

    Sutti, Salvatore; Rigamonti, Cristina; Vidali, Matteo; Albano, Emanuele

    2014-01-01

    Autoimmune reactions involving cytochrome P4502E1 (CYP2E1) are a feature of idiosyncratic liver injury induced by halogenated hydrocarbons and isoniazid, but are also detectable in about one third of the patients with advanced alcoholic liver disease (ALD) and chronic hepatitis C (CHC). In these latter the presence of anti-CYP2E1 auto-antibodies is an independent predictor of extensive necro-inflammation and fibrosis and worsens the recurrence of hepatitis following liver transplantation, indicating that CYP2E1-directed autoimmunity can contribute to hepatic injury. The molecular characterization of the antigens recognized by anti-CYP2E1 auto-antibodies in ALD and CHC has shown that the targeted conformational epitopes are located in close proximity on the molecular surface. Furthermore, these epitopes can be recognized on CYP2E1 expressed on hepatocyte plasma membranes where they can trigger antibody-mediated cytotoxicity. This does not exclude that T cell-mediated responses against CYP2E1 might also be involved in causing hepatocyte damage. CYP2E1 structural modifications by reactive metabolites and molecular mimicry represent important factors in the breaking of self-tolerance against CYP2E1 in, respectively, ALD and CHC. However, genetic or acquired interferences with the mechanisms controlling the homeostasis of the immune system are also likely to contribute. More studies are needed to better characterize the impact of anti-CYP2E1 autoimmunity in liver diseases particularly in relation to the fact that common metabolic alterations such as obesity and diabetes stimulates hepatic CYP2E1 expression. PMID:25462068

  14. New insights into FoxE1 functions: identification of direct FoxE1 targets in thyroid cells.

    Directory of Open Access Journals (Sweden)

    Lara P Fernández

    Full Text Available BACKGROUND: FoxE1 is a thyroid-specific forkhead transcription factor essential for thyroid gland development, as well as for the maintenance of the thyroid differentiated state in adults. FoxE1 recognizes and binds to a short DNA sequence present in thyroglobulin (Tg and thyroperoxidase (Tpo promoters, but FoxE1 binding to regulatory regions other than Tg and Tpo promoters remains almost unexplored. Improving knowledge of the regulatory functions of FoxE1 is necessary to clarify its role in endocrine syndromes and cancer susceptibility. METHODOLOGY/PRINCIPAL FINDING: In order to further investigate downstream FoxE1 targets, we performed a genome-wide expression screening after knocking-down FoxE1 and obtained new insights into FoxE1 transcriptional networks in thyroid follicular cells. After validation, we confirmed Adamts9, Cdh1, Duox2 and S100a4 as upregulated genes and Casp4, Creld2, Dusp5, Etv5, Hsp5a, Nr4a2 and Tm4sf1 as downregulated genes when FoxE1 was silenced. In promoter regions of putative FoxE1-regulated genes and also in the promoters of the classical thyroid genes Nis, Pax8 and Titf1, we performed an in silico search of the FoxE1 binding motif that was in close proximity to the NF1/CTF binding sequence, as previously described for other forkhead factors. Using chromatin immunoprecipitation we detected specific in vivo FoxE1 binding to novel regulatory regions in two relevant thyroid genes, Nis and Duox2. Moreover, we demonstrated simultaneous binding of FoxE1 and NF1/CTF to the Nis upstream enhancer region, as well as a clear functional activation of the Nis promoter by both transcription factors. CONCLUSIONS/SIGNIFICANCE: In search for potential downstream mediators of FoxE1 function in thyroid cells, we identified two novel direct FoxE1 target genes. To our knowledge, this is the first evidence regarding the implication of Nis and Duox2 in executing the transcriptional program triggered by FoxE1. Furthermore, this study points

  15. 26 CFR 1.514(e)-1 - Allocation rules.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 7 2010-04-01 2010-04-01 true Allocation rules. 1.514(e)-1 Section 1.514(e)-1... Allocation rules. Where only a portion of property is debt-financed property, proper allocation of the basis...)(iii) of § 1.514(b)-1 for illustrations of proper allocation....

  16. 26 CFR 1.642(e)-1 - Depreciation and depletion.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 8 2010-04-01 2010-04-01 false Depreciation and depletion. 1.642(e)-1 Section 1... (CONTINUED) INCOME TAXES Estates, Trusts, and Beneficiaries § 1.642(e)-1 Depreciation and depletion. An estate or trust is allowed the deductions for depreciation and depletion, but only to the extent...

  17. CYP2E1 Metabolism of Styrene Involves Allostery

    Science.gov (United States)

    Hartman, Jessica H.; Boysen, Gunnar

    2012-01-01

    We are the first to report allosterism during styrene oxidation by recombinant CYP2E1 and human liver microsomes. At low styrene concentrations, oxidation is inefficient because of weak binding to CYP2E1 (Ks = 830 μM). A second styrene molecule then binds CYP2E1 with higher affinity (Kss = 110 μM) and significantly improves oxidation to achieve a kcat of 6.3 nmol · min−1 · nmol CYP2E1−1. The transition between these metabolic cycles coincides with reported styrene concentrations in blood from exposed workers; thus, this CYP2E1 mechanism may be relevant in vivo. Scaled modeling of the in vitro-positive allosteric mechanism for styrene metabolism to its in vivo clearance led to significant deviations from the traditional model based on Michaelis-Menten kinetics. Low styrene levels were notably much less toxic than generally assumed. We interrogated the allosteric mechanism using the CYP2E1-specific inhibitor and drug 4-methylpyrazole, which we have shown binds two CYP2E1 sites. From the current studies, styrene was a positive allosteric effector on 4-methylpyrazole binding, based on a 10-fold increase in 4-methylpyrazole binding affinity from Ki 0.51 to Ksi 0.043 μM. The inhibitor was a negative allosteric effector on styrene oxidation, because kcat decreased 6-fold to 0.98 nmol · min−1 · nmol CYP2E1−1. Consequently, mixtures of styrene and other molecules can induce allosteric effects on binding and metabolism by CYP2E1 and thus mitigate the efficiency of their metabolism and corresponding effects on human health. Taken together, our elucidation of mechanisms for these allosteric reactions provides a powerful tool for further investigating the complexities of CYP2E1 metabolism of drugs and pollutants. PMID:22807108

  18. A distance regular graph of type E1 Ed

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    In this note, the distance regular graph of type E1 Ed is considered and some characterization of the type graph is given. The results generalize the characterization of tight distance regular graphs.

  19. Oncolytic Replication of E1b-Deleted Adenoviruses

    Directory of Open Access Journals (Sweden)

    Pei-Hsin Cheng

    2015-11-01

    Full Text Available Various viruses have been studied and developed for oncolytic virotherapies. In virotherapy, a relatively small amount of viruses used in an intratumoral injection preferentially replicate in and lyse cancer cells, leading to the release of amplified viral particles that spread the infection to the surrounding tumor cells and reduce the tumor mass. Adenoviruses (Ads are most commonly used for oncolytic virotherapy due to their infection efficacy, high titer production, safety, easy genetic modification, and well-studied replication characteristics. Ads with deletion of E1b55K preferentially replicate in and destroy cancer cells and have been used in multiple clinical trials. H101, one of the E1b55K-deleted Ads, has been used for the treatment of late-stage cancers as the first approved virotherapy agent. However, the mechanism of selective replication of E1b-deleted Ads in cancer cells is still not well characterized. This review will focus on three potential molecular mechanisms of oncolytic replication of E1b55K-deleted Ads. These mechanisms are based upon the functions of the viral E1B55K protein that are associated with p53 inhibition, late viralmRNAexport, and cell cycle disruption.

  20. Primary scientific results of Chang’E-1 lunar mission

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The strategic plan for the development of the unmanned Chinese Lunar Exploration Program is characterized by three distinct stages: "orbiting around", "landing on" and "returning from" the Moon. The first Chinese lunar probe, Chang’E-1, which was successfully launched on October 24th, 2007 at Xichang Satellite Launch Center, and guided to crash on the Moon on March 1st, 2009, at 52.36°E, 1.50°S, in the north of Mare Fecunditatis, is the first step towards the "orbiting around" stage. The Chang’E-1 mission lasted 495 days, exceeding the expected life-span by about four months. A total of 1.37 TB raw data was received from Chang’E-1. It was then processed into 4 TB scientific data products at various levels. Many scientific results have been obtained by analyzing these data, including especially the "global lunar image from the first Chinese lunar explora- tion mission". All scientific goals of Chang’E-1 have been achieved. It provides much useful materials for further advances of lunar sciences and planetary chemistry. Meanwhile, these results will serve as a firm basis for future Chinese lunar missions.

  1. Performance Analysis of Two Ethernet over E1 Schemes

    Institute of Scientific and Technical Information of China (English)

    CHEN Wentao; JIN Depeng; ZENG Lieguang

    2007-01-01

    The Ethernet over E1 approach,which takes advantage of widely deployed telecom networks,is an efficient and economical way to interconnect two Ethernets in different regions.Two Ethernet over E1 schemes, namely a byte granularity scheme and a frame granularity scheme are discussed. The byte granularity scheme partitions Ethernet frames into several pieces for transmission and has a strict requirement on the maximum delay difference of multiple E1 links. To solve this problem, the newly proposed frame granularity scheme transmits separately each frame through E1 links without any partitioning. The architecture designs of both schemes are presented. This paper evaluates the throughput and delay performances of both schemes, both analytically from results calculated from delay models and using test results from field programmable gate array (FPGA) implementation. Although the frame granularity scheme has a slightly worse delay performance, it has a higher throughput, and is the only choice able to overcome large delay differences of the E1 links.

  2. Chang'e-1 Satellite Completed Its Preset Objectives

    Institute of Scientific and Technical Information of China (English)

    He Ying

    2008-01-01

    @@ By October 23, Chang'e-1 satellite with a one-year design lifetime has been operating in lunar orbit for one year, completed more than 4000 orbits, covering the entire moon 12 times. The satellite's platform works normally at present and all systems and equipment onboard work in their main mode. The satellite has obtained a large quantity of scientific data and achieved the preset objectives of precise orbit maneuver, successful moon orbiting, effective exploration and oneyear lifetime. The Chang'e-1 mission is a complete success.

  3. 26 CFR 31.3302(e)-1 - Successor employer.

    Science.gov (United States)

    2010-04-01

    ... TAXES AND COLLECTION OF INCOME TAX AT SOURCE EMPLOYMENT TAXES AND COLLECTION OF INCOME TAX AT SOURCE Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3302(e)-1 Successor employer. (a) In general. In addition to the credits against the tax allowable under section 3302(a) and...

  4. Osteogenic gene expression of murine osteoblastic (MC3T3-E1) cells under cyclic tension

    Science.gov (United States)

    Kao, C. T.; Chen, C. C.; Cheong, U.-I.; Liu, S. L.; Huang, T. H.

    2014-08-01

    Low-level laser therapy (LLLT) can promote cell proliferation. The remodeling ability of the tension side of orthodontic teeth affects post-orthodontic stability. The purpose of the present study was to investigate the osteogenic effects of LLLT on osteoblast-like cells treated with a simulated tension system that provides a mechanical tension regimen. Murine osteoblastic (MC3T3-E1) cells were cultured in a Flexcell strain unit with programmed loads of 12% elongation at a frequency of 0.5 Hz for 24 and 48 h. The cultured cells were treated with a low-level diode laser using powers of 5 J and 10 J. The proliferation of MC3T3-E1 cells was determined using the Alamar Blue assay. The expression of osteogenic genes (type I collagen (Col-1), osteopontin (OPN), osteocalcin (OC), osteoprotegerin (OPG), receptor activator of nuclear factor kappa B ligand (RANKL), bone morphologic protein (BMP-2), and bone morphologic protein (BMP-4)) in MC3T3-E1 cells was analyzed using reverse transcription polymerase chain reaction (RT-PCR). The data were analyzed using one-way analysis of variance. The proliferation rate of tension-cultured MC3T3-E1 cells under 5 J and 10 J LLLT increased compared with that of the control group (p < 0.05). Prominent mineralization of the MC3T3-E1 cells was visible using a von Kossa stain in the 5 J LLLT group. Osteogenic genes (Col-1, OC, OPG and BMP-2) were significantly expressed in the MC3T3-E1 cells treated with 5 J and 10 J LLLT (p < 0.05). LLLT in tension-cultured MC3T3-E1 cells showed synergistic osteogenic effects, including increases in cell proliferation and Col-1, OPN, OC, OPG and BMP-2 gene expression. LLLT might be beneficial for bone remodeling on the tension side of orthodontics.

  5. The hepatitis C virus E1 glycoprotein undergoes productive folding but accelerated degradation when expressed as an individual subunit in CHO cells.

    Directory of Open Access Journals (Sweden)

    Valentina Botti

    Full Text Available Hepatitis C Virus E1E2 heterodimers are components of the viral spike. Although there is a general agreement on the necessity of the co-expression of both E1 and E2 on a single coding unit for their productive folding and assembly, in a previous study using an in vitro system we obtained strong indications that E1 can achieve folding in absence of E2. Here, we have studied the folding pathway of unescorted E1 from stably expressing CHO cells, compared to the folding observed in presence of the E2 protein. A DTT-resistant conformation is achieved by E1 in both situations, consistent with the presence of an E2-independent oxidative pathway. However, while the E1E2 heterodimer is stable inside cells, E1 expressed alone is degraded within a few hours. On the other hand, the oxidation and stability of individually expressed E2 subunits is dependent on E1 co-expression. These data are consistent with E1 and E2 assisting each other for correct folding via different mechanisms: E2 assists E1 by stabilizing a semi-native conformation meanwhile E1 drives E2 towards a productive folding pathway.

  6. Phenomenology of heavy quarkonium radiative E1 transitions

    Energy Technology Data Exchange (ETDEWEB)

    Martinez, Hector E. [Physik-Department, Technische Universität München, James-Franck-Str. 1, 85748 Garching (Germany)

    2016-01-22

    We present preliminary results of the evaluation of the next-to-leading-order (NLO) relativistic corrections to the heavy quarkonium electric dipole transition (E1) rate. In our evaluation we use the quark-antiquark potential up to 1/m{sup 2} corrections that includes the effective string theory expression for the long range, a review on the method to construct this potential is given. Our results compare favorable with the experiments and may provide predictions for the rates for which no experimental data is yet available.

  7. The chang’E-1 topographic atlas of the Moon

    CERN Document Server

    Li, Chunlai; Mu, Lingli; Ren, Xin; Zuo, Wei

    2016-01-01

    This atlas is based on the lunar global Digital Elevation Models (DEM) of Chang'E-1 (CE-1), and presents CCD stereo image data with digital photogrammetry. The spatial resolution of the DEM in this atlas is 500m, with horizontal accuracy of 192m and vertical accuracy of 120m. Color-shaded relief maps with contour lines are used to show the lunar topographical characteristics. The topographical data gathered by CE-1 can provide fundamental information for the study of lunar topographical, morphological and geological structures, as well as for lunar evolution research.

  8. The role of cyclin E1 in hepatocellular carcinoma

    OpenAIRE

    Chan, Yan-yan; 陳茵茵

    2014-01-01

    Hepatocellular carcinoma (HCC) accounts for 70-85% of liver cancer, which is the sixth most common cancer in the world. Prognosis of HCC is dismal with little chance of complete recovery after diagnosis. It is of essence to discover the key molecules involved in the tumor progression. This could help earlier detection of HCC and establish targeted molecular therapies. Cyclin E1 (CCNE1) is a cyclin molecule responsible for the transition from G1 to S phase of the cell cycle and is often dysreg...

  9. Crossover Analysis of CHANG'E-1 Laser Altimeter Data

    Science.gov (United States)

    Hu, W.; Yue, Z.; Di, K.

    2011-08-01

    This paper presents a preliminary result of crossover analysis and adjustment of Chang'E-1(CE-1) Laser Altimeter (LAM) data of the Moon for global and regional mapping applications. During the operation of Chang'E-1 from November 28, 2007 to December 4, 2008, the laser altimeter acquired 1400 orbital profiles with about 9.12 million altimetric points. In our experiment, we derived more than 1.38 million crossovers from 1395 ground tracks covering the entire lunar surface after eliminating outliers of orbits and altimetric points. A method of least-squares crossover adjustment with a series of basis functions of time (trigonometric functions and polynomials) is developed to reconcile the LAM data by minimizing the crossover residuals globally. The normal equations are very large but sparse; therefore they are stored and solved using sparse matrix technique. In a test area (0°N~60°N, 50°W~0°W), the crossover residuals are reduced from 62.1m to 32.8m, and the quality of the DEM generated from the adjusted LAM data is improved accordingly. We will optimize the method for the global adjustment to generate a high precision consistent global DEM, which can be used as absolute control for lunar mapping with orbital images.

  10. Low Cost Network Emulator with Ethernet and E1 Interfaces

    Directory of Open Access Journals (Sweden)

    Zbynek Kocur

    2010-01-01

    Full Text Available Contemporary Next Generation Networks (NGN are mainly built on the Internet Protocol (IP and Ethernet. Major challenge for emerging types of wired and wireless IP-based networks is to provide an adequate Quality of Service (QoS for different services. The quality of evaluation requires a detailed knowledge of the performance requirements for particular services and applications. The paper is primarily oriented to the end-to-end testing for the Ethernet-based terminal equipment. The low cost Ethernet network emulator was developed on the Department of Telecommunication Technology of the Faculty of Electrical Engineering of the Czech Technical University in Prague. The extension for emulation network with the E1 interfaces and TDM over IP transmission can be used with external converters.

  11. Subcellular Localization of Rat CYP2E1 Impacts Metabolic Efficiency toward Common Substrates

    OpenAIRE

    2015-01-01

    Cytochrome P450 2E1 (CYP2E1) detoxifies or bioactivates many low molecular-weight compounds. Most knowledge about CYP2E1 activity relies on studies of the enzyme localized to endoplasmic reticulum (erCYP2E1); however, CYP2E1 undergoes transport to mitochondria (mtCYP2E1) and becomes metabolically active. We report the first comparison of in vitro steady-state kinetic profiles for erCYP2E1 and mtCYP2E1 oxidation of probe substrate 4-nitrophenol and pollutants styrene and aniline using subcellu...

  12. Data of evolutionary structure change: 1E1QA-2HLDT [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1QA-2HLDT 1E1Q 2HLD A T DLEETGRVLSIGDGIARVHGLRNVQAEEMVEFSSGLKGM...VDALVPIGRGQRELIIGDRQTGKTAVALDTILNQKRWNNGSDESKKLYCVYVAVGQKRSTVAQLVQTLEQHDAMKYSIIVAATASEAAPLQYLAPFTAASIGEWFRDN...bID> T 2HLDT ALKQVAGS

  13. Data of evolutionary structure change: 1E1RC-2HLDT [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1RC-2HLDT 1E1R 2HLD C T ADTSVDLEETGRVLSIGDGIARVHGLRNVQAEEMVEFSS...VQTGLKAVDALVPIGRGQRELIIGDRQTGKTAVALDTILNQKRWNNGSDESKKLYCVYVAVGQKRSTVAQLVQTLEQHDAMKYSIIVAATASEAAPLQYLAPFTAASI... 2HLD T 2HLDT AAFAQ--

  14. Data of evolutionary structure change: 1E1RA-2HLDT [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1RA-2HLDT 1E1R 2HLD A T DLEETGRVLSIGDGIARVHGLRNVQAEEMVEFSSGLKGM...VDALVPIGRGQRELIIGDRQTGKTAVALDTILNQKRWNNGSDESKKLYCVYVAVGQKRSTVAQLVQTLEQHDAMKYSIIVAATASEAAPLQYLAPFTAASIGEWFRDN...bChain>T 2HLDT ALKQVAGSLKLFL

  15. N′-[(1E-1-(4-Chlorophenylethylidene]formohydrazide

    Directory of Open Access Journals (Sweden)

    Zahid Shafiq

    2009-10-01

    Full Text Available The structure of the title compound, C9H9ClN2O, consists of centrosymmetric dimers due to intermolecular N—H...O hydrogen bonding, forming R22(8 ring motifs. The dihedral angle between the p-chlorophenyl unit and the remaining heavy-atom group is 6.77 (17°.

  16. Estrogen sulfotransferase/SULT1E1 promotes human adipogenesis.

    Science.gov (United States)

    Ihunnah, Chibueze A; Wada, Taira; Philips, Brian J; Ravuri, Sudheer K; Gibbs, Robert B; Kirisci, Levent; Rubin, J Peter; Marra, Kacey G; Xie, Wen

    2014-05-01

    Estrogen sulfotransferase (EST/SULT1E1) is known to catalyze the sulfoconjugation and deactivation of estrogens. The goal of this study is to determine whether and how EST plays a role in human adipogenesis. By using human primary adipose-derived stem cells (ASCs) and whole-fat tissues from the abdominal subcutaneous fat of obese and nonobese subjects, we showed that the expression of EST was low in preadipocytes but increased upon differentiation. Overexpression and knockdown of EST in ASCs promoted and inhibited differentiation, respectively. The proadipogenic activity of EST in humans was opposite to the antiadipogenic effect of the same enzyme in rodents. Mechanistically, EST promoted adipogenesis by deactivating estrogens. The proadipogenic effect of EST can be recapitulated by using an estrogen receptor (ER) antagonist or ERα knockdown. In contrast, activation of ER in ASCs inhibited adipogenesis by decreasing the recruitment of the adipogenic peroxisome proliferator-activated receptor γ (PPARγ) onto its target gene promoters, whereas ER antagonism increased the recruitment of PPARγ to its target gene promoters. Linear regression analysis revealed a positive correlation between the expression of EST and body mass index (BMI), as well as a negative correlation between ERα expression and BMI. We conclude that EST is a proadipogenic factor which may serve as a druggable target to inhibit the turnover and accumulation of adipocytes in obese patients.

  17. E1 and M1 strength functions at low energy

    Directory of Open Access Journals (Sweden)

    Schwengner Ronald

    2017-01-01

    Full Text Available We report photon-scattering experiments using bremsstrahlung at the γELBE facility of Helmholtz-Zentrum Dresden-Rossendorf and using quasi-monoenergetic, polarized γ beams at the HIγS facility of the Triangle Universities Nuclear Laboratory in Durham. To deduce the photoabsorption cross sections at high excitation energy and high level density, unresolved strength in the quasicontinuum of nuclear states has been taken into account. In the analysis of the spectra measured by using bremsstrahlung at γELBE, we perform simulations of statistical γ-ray cascades using the code γDEX to estimate intensities of inelastic transitions to low-lying excited states. Simulated average branching ratios are compared with model-independent branching ratios obtained from spectra measured by using monoenergetic γ beams at HIγS. E1 strength in the energy region of the pygmy dipole resonance is discussed in nuclei around mass 90 and in xenon isotopes. M1 strength in the region of the spin-flip resonance is also considered for xenon isotopes. The dipole strength function of 74Ge deduced from γELBE experiments is compared with the one obtained from experiments at the Oslo Cyclotron Laboratory. The low-energy upbend seen in the Oslo data is interpreted as M1 strength on the basis of shell-model calculations.

  18. Data of evolutionary structure change: 1E1QC-2HLDA [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1QC-2HLDA 1E1Q 2HLD C A ADTSVDLEETGRVLSIGDGIARVHGLRNVQAEEMVEFSS...ELFYKGIRPAINVGLSVSRVGSAAQTRAMKQVAGTMKLELAQYREVAAFAQFGSDLDAATQQLLSRGVRLTELLKQGQYSPMAIEEQVAVIYAGVRGYLDKLEPSKIT...RPAINVGLSVSRVGSAAQVKALKQVAGSLKLFLAQYREVAAFAQS--DLDASTKQTLVRGERLTQLLKQNQYSPLATEEQV...> 0 2HLD A 2HLDA AFAQS--DLDA...ain> 1E1Q C 1E1QC AFAQFGSDLDA

  19. Bis[(E-1-(3,4-dichlorobenzylideneamino-4-methylpyridinium] bis(maleonitriledithiolatonickelate(II

    Directory of Open Access Journals (Sweden)

    Shao-Ming Zhang

    2009-02-01

    Full Text Available The asymmetric unit of the title compound, (C13H11Cl2N22[Ni(C4N2S22], contains one-half of a centrosymmetric [Ni(mnt2] anion (where mnt is maleonitriledithiolate or 1,2-dicyano-1,2-ethylenedithiolate and an (E-1-(3,4-dichlorobenzylideneamino-4-methylpyridinium cation. In the anion, the coordination around the Ni atom is a distorted square. In the cation, the aromatic rings are oriented at a dihedral angle of 7.81 (3°. In the crystal structure, intermolecular C—H...N hydrogen bonds link the cations and anions. π–π Contacts between the nickel dithiolene and pyridine rings and between the benzene and pyridine rings, [centroid–centroid distances = 3.682 (3 and 3.643 (3 Å, respectively] may further stabilize the structure.

  20. Metabolic inactivation of resolvin E1 and stabilization of its anti-inflammatory actions.

    Science.gov (United States)

    Arita, Makoto; Oh, Sungwhan F; Chonan, Tomomichi; Hong, Song; Elangovan, Siva; Sun, Yee-Ping; Uddin, Jasim; Petasis, Nicos A; Serhan, Charles N

    2006-08-11

    The resolvins (Rv) are lipid mediators derived from omega-3 polyunsaturated fatty acids that act within a local inflammatory milieu to stop leukocyte recruitment and promote resolution. Resolvin E1 (RvE1; (5S,12R,18R)-trihydroxy-6Z,8E,10E,14Z,16E-eicosapentaenoic acid) is an oxygenase product derived from omega-3 eicosapentaenoic acid that displays potent anti-inflammation/pro-resolution actions in vivo. Here, we determined whether oxidoreductase enzymes catalyze the conversion of RvE1 and assessed the biological activity of the RvE1 metabolite. With NAD+ as a cofactor, recombinant 15-hydroxyprostaglandin dehydrogenase acted as an 18-hydroxyl dehydrogenase to form 18-oxo-RvE1. In the murine lung, dehydrogenation of the hydroxyl group at carbon 18 position to form 18-oxo-RvE1 represented the major initial metabolic route for RvE1. At a concentration where RvE1 potently reduced polymorphonuclear leukocyte (PMN) recruitment in zymosan-induced peritonitis, 18-oxo-RvE1 was devoid of activity. In human neutrophils, carbon 20 hydroxylation of RvE1 was the main route of conversion. An RvE1 analog, i.e. 19-(p-fluorophenoxy)-RvE1, was synthesized that resisted rapid metabolic inactivation and proved to retain biological activity reducing PMN infiltration and pro-inflammatory cytokine/chemokine production in vivo. These results established the structure of a novel RvE1 initial metabolite, indicating that conversion of RvE1 to the oxo product represents a mode of RvE1 inactivation. Moreover, the designed RvE1 analog, which resisted further metabolism/inactivation, could be a useful tool to evaluate the actions of RvE1 in complex disease models.

  1. Data of evolutionary structure change: 1E1QC-2HLDT [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1QC-2HLDT 1E1Q 2HLD C T ADTSVDLEETGRVLSIGDGIARVHGLRNVQAEEMVEFSS...VQTGLKAVDALVPIGRGQRELIIGDRQTGKTAVALDTILNQKRWNNGSDESKKLYCVYVAVGQKRSTVAQLVQTLEQHDAMKYSIIVAATASEAAPLQYLAPFTAASI...yChain> 2HLD T 2HLDT VAAFA---QDLD

  2. Data of evolutionary structure change: 1E1QB-2HLDT [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1QB-2HLDT 1E1Q 2HLD B T DLEETGRVLSIGDGIARVHGLRNVQAEEMVEFSSGLKGM...SDRLVKEGELVKRTGNIVDVPVGPGLLGRVVDALGNPIDGKGPIDAAGRSRAQVKAPGILPRRSVHEPVQTGLKAVDALVPIGRGQRELIIGDRQTGKTAVALDT...dbChain> 2HLDT ALKQVAGSLKLFL 2HLD T 2HLDT VAAFAQDLDAST

  3. Data of evolutionary structure change: 1E1RB-2HLDT [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1RB-2HLDT 1E1R 2HLD B T DLEETGRVLSIGDGIARVHGLRNVQAEEMVEFSSGLKGM...SDRLVKEGELVKRTGNIVDVPVGPGLLGRVVDALGNPIDGKGPIDAAGRSRAQVKAPGILPRRSVHEPVQTGLKAVDALVPIGRGQRELIIGDRQTGKTAVALDT...T 2HLDT ALKQVAGSLKLFL2HLD T 2HLDT VAAFAQDLDAST

  4. A note on inextensible flows of partially and pseudo null curves in E_1^4

    OpenAIRE

    Yuzbasi, Zuhal Kucukarslan; Bektas, Mehmet

    2013-01-01

    In this paper, we study inextensible flows of partially null and pseudo null curves in E_1^4. We give neccessary and sufficent conditions for inextensible flows of partially null and pseudo null curves in E_1^4

  5. Data of evolutionary structure change: 1E1RC-2HLDA [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1E1RC-2HLDA 1E1R 2HLD C A ADTSVDLEETGRVLSIGDGIARVHGLRNVQAEEMVEFSS...ELFYKGIRPAINVGLSVSRVGSAAQTRAMKQVAGTMKLELAQYREVAAFAQFGSDLDAATQQLLSRGVRLTELLKQGQYSPMAIEEQVAVIYAGVRGYLDKLEPSKIT...RPAINVGLSVSRVGSAAQVKALKQVAGSLKLFLAQYREVAAFAQS--DLDASTKQTLVRGERLTQLLKQNQYSPLATEEQV...fEVID> 0 2HLD A 2HLDA AFAQS--DLDAS GGG -- HH

  6. Cell-based analysis of Chikungunya virus E1 protein in membrane fusion

    Directory of Open Access Journals (Sweden)

    Kuo Szu-Cheng

    2012-04-01

    Full Text Available Abstract Background Chikungunya fever is a pandemic disease caused by the mosquito-borne Chikungunya virus (CHIKV. E1 glycoprotein mediation of viral membrane fusion during CHIKV infection is a crucial step in the release of viral genome into the host cytoplasm for replication. How the E1 structure determines membrane fusion and whether other CHIKV structural proteins participate in E1 fusion activity remain largely unexplored. Methods A bicistronic baculovirus expression system to produce recombinant baculoviruses for cell-based assay was used. Sf21 insect cells infected by recombinant baculoviruses bearing wild type or single-amino-acid substitution of CHIKV E1 and EGFP (enhanced green fluorescence protein were employed to investigate the roles of four E1 amino acid residues (G91, V178, A226, and H230 in membrane fusion activity. Results Western blot analysis revealed that the E1 expression level and surface features in wild type and mutant substituted cells were similar. However, cell fusion assay found that those cells infected by CHIKV E1-H230A mutant baculovirus showed little fusion activity, and those bearing CHIKV E1-G91D mutant completely lost the ability to induce cell-cell fusion. Cells infected by recombinant baculoviruses of CHIKV E1-A226V and E1-V178A mutants exhibited the same membrane fusion capability as wild type. Although the E1 expression level of cells bearing monomeric-E1-based constructs (expressing E1 only was greater than that of cells bearing 26S-based constructs (expressing all structural proteins, the sizes of syncytial cells induced by infection of baculoviruses containing 26S-based constructs were larger than those from infections having monomeric-E1 constructs, suggesting that other viral structure proteins participate or regulate E1 fusion activity. Furthermore, membrane fusion in cells infected by baculovirus bearing the A226V mutation constructs exhibited increased cholesterol-dependences and lower pH thresholds

  7. Absence of NR2E1 mutations in patients with aniridia

    DEFF Research Database (Denmark)

    Corso-Díaz, Ximena; Borrie, Adrienne E; Bonaguro, Russell;

    2012-01-01

    Nuclear receptor 2E1 (NR2E1) is a transcription factor with many roles during eye development and thus may be responsible for the occurrence of certain congenital eye disorders in humans. To test this hypothesis, we screened NR2E1 for candidate mutations in patients with aniridia and other congen...

  8. Infection with E1B-mutant adenovirus stabilizes p53 but blocks p53 acetylation and activity through E1A

    DEFF Research Database (Denmark)

    Savelyeva, I.; Dobbelstein, M.

    2011-01-01

    Wild-type adenovirus type 5 eliminates p53 through the E1B-55 kDa and E4-34 kDa gene products. Deletion or mutation of E1B-55 kDa has long been thought to confer p53-selective replication of oncolytic viruses. We show here that infection with E1B-defective adenovirus mutants induces massive...... accumulation of p53, without obvious defects in p53 localization, phosphorylation, conformation and oligomerization. Nonetheless, p53 completely failed to induce its target genes in this scenario, for example, p21/CDKN1A, Mdm2 and PUMA. Two regions of the E1A gene products independently contributed...... acetylation in infected cells. Mutating either of these E1A regions, in addition to E1B, partially restored p21 mRNA levels. Our findings argue that adenovirus attenuates p53-mediated p21 induction, through at least two E1B-independent mechanisms. Other virus species and cancer cells may employ analogous...

  9. Structural models of the membrane anchors of envelope glycoproteins E1 and E2 from pestiviruses

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jimin, E-mail: jimin.wang@yale.edu; Li, Yue; Modis, Yorgo, E-mail: yorgo.modis@yale.edu

    2014-04-15

    The membrane anchors of viral envelope proteins play essential roles in cell entry. Recent crystal structures of the ectodomain of envelope protein E2 from a pestivirus suggest that E2 belongs to a novel structural class of membrane fusion machinery. Based on geometric constraints from the E2 structures, we generated atomic models of the E1 and E2 membrane anchors using computational approaches. The E1 anchor contains two amphipathic perimembrane helices and one transmembrane helix; the E2 anchor contains a short helical hairpin stabilized in the membrane by an arginine residue, similar to flaviviruses. A pair of histidine residues in the E2 ectodomain may participate in pH sensing. The proposed atomic models point to Cys987 in E2 as the site of disulfide bond linkage with E1 to form E1–E2 heterodimers. The membrane anchor models provide structural constraints for the disulfide bonding pattern and overall backbone conformation of the E1 ectodomain. - Highlights: • Structures of pestivirus E2 proteins impose constraints on E1, E2 membrane anchors. • Atomic models of the E1 and E2 membrane anchors were generated in silico. • A “snorkeling” arginine completes the short helical hairpin in the E2 membrane anchor. • Roles in pH sensing and E1–E2 disulfide bond formation are proposed for E1 residues. • Implications for E1 ectodomain structure and disulfide bonding pattern are discussed.

  10. [Induction of rat hepatic CYP2E1 expression by arecoline in vivo].

    Science.gov (United States)

    Huang, Xiang-tao; Xiao, Run-mei; Wang, Ming-feng; Wang, Jun-jun; Chen, Yong

    2016-01-01

    The regulation mechanism of arecoline on rat hepatic CYP2E1 was studied in vivo. After oral administration of arecoline hydrobromide (AH; 4, 20 and 100 mg x kg(-1) x d(-1)) to rats for one week, the hepatic CYP2E1 mRNA level remained unchanged, but the hepatic CYP2E1 protein content was dose-dependently increased. Additionally, although the hepatic CYP2E1 activity was induced by AH treatment, the induction was attenuated with the increase in dosage. The results indicate that the effect of arecoline on rat hepaticdoes not involve transcriptional activation of the gene, but largely involves the stabilization of CYP2E1 protein against degradation or increased efficiency of CYP2E1 mRNA translation, and additionally involve the post- ranslational modification of CYP2E1 protein. Furthermore, the CYP2E1 response is fairly equal among the different species, the induction of rat hepatic CYP2E1 by arecoline suggests that there is a risk of metabolic interaction among the substrate drugs of CYP2E1 in betel-quid use human.

  11. Improved Plant-based Production of E1 endoglucanase Using Potato: Expression Optimization and Tissue Targeting

    Energy Technology Data Exchange (ETDEWEB)

    Dai, Ziyu [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Hooker, Brian S. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Anderson, Daniel B. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Thomas, Steven R. [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2000-06-01

    Optimization of Acidothermus cellulolyticus endoglucanase (E1) gene expression in transgenic potato (Solanum tuberosum L.) was examined in this study, where the E1 coding sequence was transcribed under control of a leaf specific promoter (tomato RbcS-3C) or the Mac promoter (a hybrid promoter of mannopine synthase promoter and cauliflower mosaic virus 35S promoter enhancer region). Average E1 activity in leaf extracts of potato transformants, in which E1 protein was targeted by a chloroplast signal peptide and an apoplast signal peptide were much higher than those by an E1 native signal peptide and a vacuole signal peptide. E1 protein accumulated up to 2.6% of total leaf soluble protein, where E1 gene was under control of the RbcS-3C promoter, alfalfa mosaic virus 5-untranslated leader, and RbcS-2A signal peptide. E1 protein production, based on average E1 activity and E1 protein accumulation in leaf extracts, is higher in potato than those measured previously in transgenic tobacco bearing the same transgene constructs. Comparisons of E1 activity, protein accumulation, and relative mRNA levels showed that E1 expression under control of tomato RbcS-3C promoter was specifically localized in leaf tissues, while E1 gene was expressed in both leaf and tuber tissues under control of Mac promoter. This suggests dual-crop applications in which potato vines serve as enzyme production `bioreactors' while tubers are preserved for culinary applications.

  12. Effect of glucocorticoid on prostaglandin E1 mediated cyclic AMP formation by vascular smooth muscle cells.

    Science.gov (United States)

    Yasunari, K; Kohno, M; Murakawa, K; Yokokawa, K; Takeda, T

    1988-12-01

    The effect of glucocorticoid on the prostaglandin E1 (PGE1)-mediated cyclic AMP (cAMP) formation by vascular smooth muscle cells (VSMC) from renal arteries (RA) was studied in rats. Dexamethasone (DEX) at concentrations ranging from 10(-10) to approximately 10(-8) mol/l dose-dependently potentiates the PGE1-mediated response. This facilitation began at 6 h and reached its maximum after 24 h of DEX administration. Aldosterone (10(-6) mol/l) did not affect the dose-response curve of PGE1. Inhibitors of protein and RNA synthesis blocked this glucocorticoid effect. The basal activity of adenylate cyclase in DEX-treated cells was twice as high as in control cells. Treatment of VSMC with DEX increased cholera toxin- and pertussis toxin-stimulated adenylate cyclase activity. DEX treatment also augments forskolin-stimulated adenylate cyclase activity. These results suggest that DEX increases PGE1-mediated cAMP formation of VSMC from RA through a mechanism that involves the induction of protein synthesis, and that the activation of the catalytic unit may play some role in this facilitating process.

  13. The Role of CYP2E1 in the Drug Metabolism or Bioactivation in the Brain

    Science.gov (United States)

    García-Suástegui, W. A.; Ramos-Chávez, L. A.; Rubio-Osornio, M.; Calvillo-Velasco, M.; Atzin-Méndez, J. A.; Guevara, J.

    2017-01-01

    Organisms have metabolic pathways that are responsible for removing toxic agents. We always associate the liver as the major organ responsible for detoxification of the body; however this process occurs in many tissues. In the same way, as in the liver, the brain expresses metabolic pathways associated with the elimination of xenobiotics. Besides the detoxifying role of CYP2E1 for compounds such as electrophilic agents, reactive oxygen species, free radical products, and the bioactivation of xenobiotics, CYP2E1 is also related in several diseases and pathophysiological conditions. In this review, we describe the presence of phase I monooxygenase CYP2E1 in regions of the brain. We also explore the conditions where protein, mRNA, and the activity of CYP2E1 are induced. Finally, we describe the relation of CYP2E1 in brain disorders, including the behavioral relations for alcohol consumption via CYP2E1 metabolism. PMID:28163821

  14. The Role of CYP2E1 in the Drug Metabolism or Bioactivation in the Brain

    Directory of Open Access Journals (Sweden)

    W. A. García-Suástegui

    2017-01-01

    Full Text Available Organisms have metabolic pathways that are responsible for removing toxic agents. We always associate the liver as the major organ responsible for detoxification of the body; however this process occurs in many tissues. In the same way, as in the liver, the brain expresses metabolic pathways associated with the elimination of xenobiotics. Besides the detoxifying role of CYP2E1 for compounds such as electrophilic agents, reactive oxygen species, free radical products, and the bioactivation of xenobiotics, CYP2E1 is also related in several diseases and pathophysiological conditions. In this review, we describe the presence of phase I monooxygenase CYP2E1 in regions of the brain. We also explore the conditions where protein, mRNA, and the activity of CYP2E1 are induced. Finally, we describe the relation of CYP2E1 in brain disorders, including the behavioral relations for alcohol consumption via CYP2E1 metabolism.

  15. Inhibitory Potency of 4-Carbon Alkanes and Alkenes toward CYP2E1 Activity

    OpenAIRE

    2014-01-01

    CYP2E1 has been implicated in the bioactivation of many small molecules into reactive metabolites which form adducts with proteins and DNA, and thus a better understanding of the molecular determinants of its selectivity are critical for accurate toxicological predictions. In this study, we determined the potency of inhibition of human CYP2E1 for various 4-carbon alkanes, alkenes and alcohols. In addition, known CYP2E1 substrates and inhibitors including 4-methylpyrazole, aniline, and dimethy...

  16. CYP2E1 mediated isoniazid-induced hepatotoxicity in rats

    Institute of Scientific and Technical Information of China (English)

    Jiang YUE; Ren-xiu PENG; Jing YANG; Rui KONG; Juan LIU

    2004-01-01

    AIM: To investigate the role of CYP2E1 in isoniazid (INH)-induced hepatotoxicity and the influence of rifampicin (RFP) on INH-induced liver injury. METHODS: Rats were treated with INH alone (100 mg/kg, ip) or co-administered with RFP (100 mg/kg, ig) for 10 d and 21 d. Hepatotoxicity was assayed by plasma enzymes (sALT, sAST) and histopathological examinations. Hepatic CYP2E1 activity was measured by aniline hydroxylase (ANH), and CYP2E1 mRNA expression was determined by RT-PCR. Plasma hydrazine concentration was determined by RP-HPLC.RESULTS: For a 10 d INH-treatment, hepatic CYP2E1 level was increased to 3.7-fold over the control; liver impairment appeared after 21 d treatment, while CYP2E1 and plasma hydrazine were, respectively, increased to 4.6-fold and 1.7-fold. However, in INH-RFP group for 10 d, CYP2E1 and plasma hydrazine were, respectively,decreased by 13 % and 18 % over INH group; similarly, hepatic injury is equal to INH group appeared after 21 d,and CYP2E1 was further decreased by 26 %. Correlation analysis showed that sALT had a positive correlation with plasma hydrazine and with CYP2E1 activity; CYP2E1 activity was also markedly correlated with plasma hydrazine.And compared with control, there is no difference in changes of CYP2E1 mRNA expression in INH and INH-RFP treatment for 21 d. CONCLUSION: The metabolite of INH, hydrazine, plays an important role in INH-induced hepatotoxicity in rats. The induction of CYP2E1 by hydrazine is involved in the hepatotoxicity of INH. RFP does not exacerbate INH-induced hepatotoxicity in short term, which relates to down-regulation of CYP2E1.

  17. 26 CFR 1.1059(e)-1 - Non-pro rata redemptions.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 11 2010-04-01 2010-04-01 true Non-pro rata redemptions. 1.1059(e)-1 Section 1... (CONTINUED) INCOME TAXES Special Rules § 1.1059(e)-1 Non-pro rata redemptions. (a) In general. Section 1059(d... 1059(e)(1). For example, if a redemption of stock is not pro rata as to all shareholders, any...

  18. Absence of NR2E1 mutations in patients with aniridia

    DEFF Research Database (Denmark)

    Corso-Díaz, Ximena; Borrie, Adrienne E; Bonaguro, Russell;

    2012-01-01

    Nuclear receptor 2E1 (NR2E1) is a transcription factor with many roles during eye development and thus may be responsible for the occurrence of certain congenital eye disorders in humans. To test this hypothesis, we screened NR2E1 for candidate mutations in patients with aniridia and other congen...... congenital ocular malformations (anterior segment dysgenesis, congenital optic nerve malformation, and microphthalmia)....

  19. CYP2E1-dependent hepatotoxicity and oxidative damage after ethanol administration in human primary hepatocytes

    Institute of Scientific and Technical Information of China (English)

    Lie-Gang Liu; Hong Yan; Ping Yao; Wen Zhang; Li-Jun Zou; Fang-Fang Song; Ke Li; Xiu-Fa Sun

    2005-01-01

    AIM: To observe the relationship between ethanol-induced oxidative damage in human primary cultured hepatocytes and cytochrome P450 2E1 (CYP2E1) activity, in order to address if inhibition of CYP2E1 could attenuate ethanol-induced cellular damage.METHODS: The dose-dependent (25-100 mmol/L) and time-dependent (0-24 h) exposures of primary human cultured hepatocytes to ethanol were carried out. CYP2E1 activity and protein expression were detected by spectrophotometer and Western blot analysis respectively.Hepatotoxicity was investigated by determination of lactate dehydrogenase (LDH) and aspartate transaminase (AST) level in hepatocyte culture supernatants, as well as the intracellular formation of malondialdehyde (MDA).RESULTS: A dose-and time-dependent response between ethanol exposure and CYP2E1 activity in human hepatocytes was demonstrated. Moreover, there was a time-dependent increase of CYP2E1 protein after 100 mmol/L ethanol exposure. Meanwhile, ethanol exposure of hepatocytes caused a time-dependent increase of ceilular MDA level, LDH, and AST activities in supernatants.Furthermore, the inhibitor of CYP2E1, diallyl sulfide (DAS) could partly attenuate the increases of MDA, LDH, and AST in human hepatocytes.CONCLUSION: A positive relationship between ethanol-induced oxidative aamage in human primary cultured hepatocytes and CYP2E1 activity was exhibited, and the inhibition of CYP2E1 could partly attenuate ethanol-induced oxidative damage.

  20. Requirements for E1A dependent transcription in the yeast Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Mymryk Joe S

    2009-04-01

    Full Text Available Abstract Background The human adenovirus type 5 early region 1A (E1A gene encodes proteins that are potent regulators of transcription. E1A does not bind DNA directly, but is recruited to target promoters by the interaction with sequence specific DNA binding proteins. In mammalian systems, E1A has been shown to contain two regions that can independently induce transcription when fused to a heterologous DNA binding domain. When expressed in Saccharomyces cerevisiae, each of these regions of E1A also acts as a strong transcriptional activator. This allows yeast to be used as a model system to study mechanisms by which E1A stimulates transcription. Results Using 81 mutant yeast strains, we have evaluated the effect of deleting components of the ADA, COMPASS, CSR, INO80, ISW1, NuA3, NuA4, Mediator, PAF, RSC, SAGA, SAS, SLIK, SWI/SNF and SWR1 transcriptional regulatory complexes on E1A dependent transcription. In addition, we examined the role of histone H2B ubiquitylation by Rad6/Bre1 on transcriptional activation. Conclusion Our analysis indicates that the two activation domains of E1A function via distinct mechanisms, identify new factors regulating E1A dependent transcription and suggest that yeast can serve as a valid model system for at least some aspects of E1A function.

  1. Human CYP2E1 mediates the formation of glycidamide from acrylamide.

    Science.gov (United States)

    Settels, Eva; Bernauer, Ulrike; Palavinskas, Richard; Klaffke, Horst S; Gundert-Remy, Ursula; Appel, Klaus E

    2008-10-01

    Regarding the cancer risk assessment of acrylamide (AA) it is of basic interest to know, as to what amount of the absorbed AA is metabolized to glycidamide (GA) in humans, compared to what has been observed in laboratory animals. GA is suspected of being the ultimate carcinogenic metabolite of AA. From experiments with CYP2E1-deficient mice it can be concluded that AA is metabolized to GA primarily by CYP2E1. We therefore examined whether CYP2E1 is involved in GA formation in non-rodent species with the focus on humans by using human CYP2E1 supersomes, marmoset and human liver microsomes and in addition, genetically engineered V79 cells expressing human CYP2E1 (V79h2E1 cells). Special emphasis was placed on the analytical detection of GA, which was performed by gas chromatography/mass spectrometry. The results show that AA is metabolized to GA in human CYP2E1 supersomes, in marmoset and human liver microsomes as well as in V79h2E1 cells. The activity of GA formation is highest in supersomes; in human liver it is somewhat higher than in marmoset liver. A monoclonal CYP2E1 human selective antibody (MAB-2E1) and diethyldithiocarbamate (DDC) were used as specific inhibitors of CYP2E1. The generation of GA could be inhibited by MAB-2E1 to about 80% in V79h2E1 cells and to about 90% in human and marmoset liver microsomes. Also DDC led to an inhibition of about 95%. In conclusion, AA is metabolized to GA by human CYP2E1. Overall, the present work describes (1) the application and refinement of a sensitive methodology in order to determine low amounts of GA, (2) the applicability of genetically modified V79 cell lines in order to investigate specific questions concerning metabolism and (3) the involvement, for the first time, of human CYP2E1 in the formation of GA from AA. Further studies will compare the activities of GA formation in genetically engineered V79 cells expressing CYP2E1 from different species.

  2. Identification and characterization of multiple conserved nuclear localization signals within adenovirus E1A

    Energy Technology Data Exchange (ETDEWEB)

    Marshall, Kris S.; Cohen, Michael J.; Fonseca, Greg J.; Todorovic, Biljana; King, Cason R. [Department of Microbiology and Immunology, Western University, London Regional Cancer Program, London, ON, Canada N6A 4L6 (Canada); Yousef, Ahmed F. [Department of Chemical and Environmental Engineering, Masdar Institute, Abu Dhabi (United Arab Emirates); Zhang, Zhiying [College of Animal Science and Technologies, Northwest A and F University, Yangling, Shaanxi 712100 (China); Mymryk, Joe S., E-mail: jmymryk@uwo.ca [Department of Microbiology and Immunology, Western University, London Regional Cancer Program, London, ON, Canada N6A 4L6 (Canada); Department of Oncology, Western University, London Regional Cancer Program, London, ON, Canada N6A 4L6 (Canada)

    2014-04-15

    The human adenovirus 5 (HAdV-5) E1A protein has a well defined canonical nuclear localization signal (NLS) located at its C-terminus. We used a genetic assay in the yeast Saccharomyces cerevisiae to demonstrate that the canonical NLS is present and functional in the E1A proteins of each of the six HAdV species. This assay also detects a previously described non-canonical NLS within conserved region 3 and a novel active NLS within the N-terminal/conserved region 1 portion of HAdV-5 E1A. These activities were also present in the E1A proteins of each of the other five HAdV species. These results demonstrate that, despite substantial differences in primary sequence, HAdV E1A proteins are remarkably consistent in that they contain one canonical and two non-canonical NLSs. By utilizing independent mechanisms, these multiple NLSs ensure nuclear localization of E1A in the infected cell. - Highlights: • HAdV E1A uses multiple mechanisms for nuclear import. • We identified an additional non-canonical NLS in the N-terminal/CR1 portion of E1A. • The new NLS does not contact importin-alpha directly. • All NLSs are functionally conserved in the E1A proteins of all 6 HAdV species.

  3. Human CYP2E1 mediates the formation of glycidamide from acrylamide

    Energy Technology Data Exchange (ETDEWEB)

    Settels, Eva; Appel, Klaus E. [Federal Institute for Risk Assessment, Center for Experimental Toxicology, Berlin (Germany); Bernauer, Ulrike; Gundert-Remy, Ursula [Federal Institute for Risk Assessment, Department of Safety of Substances and Preparations, Berlin (Germany); Palavinskas, Richard; Klaffke, Horst S. [Federal Institute for Risk Assessment, Center for Analytical Chemistry, Berlin (Germany)

    2008-10-15

    Regarding the cancer risk assessment of acrylamide (AA) it is of basic interest to know, as to what amount of the absorbed AA is metabolized to glycidamide (GA) in humans, compared to what has been observed in laboratory animals. GA is suspected of being the ultimate carcinogenic metabolite of AA. From experiments with CYP2E1-deficient mice it can be concluded that AA is metabolized to GA primarily by CYP2E1. We therefore examined whether CYP2E1 is involved in GA formation in non-rodent species with the focus on humans by using human CYP2E1 supersomes trademark, marmoset and human liver microsomes and in addition, genetically engineered V79 cells expressing human CYP2E1 (V79h2E1 cells). Special emphasis was placed on the analytical detection of GA, which was performed by gas chromatography/mass spectrometry. The results show that AA is metabolized to GA in human CYP2E1 supersomes trademark, in marmoset and human liver microsomes as well as in V79h2E1 cells. The activity of GA formation is highest in supersomes trademark; in human liver it is somewhat higher than in marmoset liver. A monoclonal CYP2E1 human selective antibody (MAB-2E1) and diethyldithiocarbamate (DDC) were used as specific inhibitors of CYP2E1. The generation of GA could be inhibited by MAB-2E1 to about 80% in V79h2E1 cells and to about 90% in human and marmoset liver microsomes. Also DDC led to an inhibition of about 95%. In conclusion, AA is metabolized to GA by human CYP2E1. Overall, the present work describes (1) the application and refinement of a sensitive methodology in order to determine low amounts of GA, (2) the applicability of genetically modified V79 cell lines in order to investigate specific questions concerning metabolism and (3) the involvement, for the first time, of human CYP2E1 in the formation of GA from AA. Further studies will compare the activities of GA formation in genetically engineered V79 cells expressing CYP2E1 from different species. (orig.)

  4. Methodology to assay CYP2E1 mixed function oxidase catalytic activity and its induction

    Directory of Open Access Journals (Sweden)

    Arthur I. Cederbaum

    2014-01-01

    Full Text Available The cytochrome P450 mixed function oxidase enzymes are the major catalysts involved in drug metabolism. There are many forms of P450. CYP2E1 metabolizes many toxicologically important compounds including ethanol and is active in generating reactive oxygen species. Since several of the contributions in the common theme series “Role of CYP2E1 and Oxidative/Nitrosative Stress in the Hepatotoxic Actions of Alcohol” discuss CYP2E1, this methodology review describes assays on how CYP2E1 catalytic activity and its induction by ethanol and other inducers can be measured using substrate probes such as the oxidation of para-nitrophenol to para-nitrocatechol and the oxidation of ethanol to acetaldehyde. Approaches to validate that a particular reaction e.g. oxidation of a drug or toxin is catalyzed by CYP2E1 or that induction of that reaction is due to induction of CYP2E1 are important and specific examples using inhibitors of CYP2E1, anti-CYP2E1 IgG or CYP2E1 knockout and knockin mice will be discussed.

  5. Crystal structures of four co-crystals of (E)-1,2-di(pyridin-4-yl)ethene with 4-alk-oxy-benzoic acids: 4-meth-oxy-benzoic acid-(E)-1,2-di(pyridin-4-yl)ethene (2/1), 4-eth-oxy-benzoic acid-(E)-1,2-di(pyridin-4-yl)ethene (2/1), 4-n-propoxybenzoic acid-(E)-1,2-di(pyridin-4-yl)ethene (2/1) and 4-n-but-oxy-benzoic acid-(E)-1,2-di(pyridin-4-yl)ethene (2/1).

    Science.gov (United States)

    Tabuchi, Yohei; Gotoh, Kazuma; Ishida, Hiroyuki

    2016-11-01

    The crystal structures of four hydrogen-bonded co-crystals of 4-alk-oxy-benzoic acid-(E)-1,2-di(pyridin-4-yl)ethene (2/1), namely, 2C8H8O3·C12H10N2, (I), 2C9H10O3·C12H10N2, (II), 2C10H12O3·C12H10N2, (III) and 2C11H14O3·C12H10N2, (IV), have been determined at 93 K. In compounds (I) and (IV), the asymmetric units are each composed of one 4-alk-oxy-benzoic acid mol-ecule and one half-mol-ecule of (E)-1,2-di(pyridin-4-yl)ethene, which lies on an inversion centre. The asymmetric unit of (II) consists of two crystallographically independent 4-eth-oxy-benzoic acid mol-ecules and one 1,2-di(pyridin-4-yl)ethene mol-ecule. Compound (III) crystallizes in a non-centrosymmetric space group (Pc) and the asymmetric unit comprises four 4-n-propoxybenzoic acid mol-ecules and two (E)-1,2-di(pyridin-4-yl)ethane mol-ecules. In each crystal, the acid and base components are linked by O-H⋯N hydrogen bonds, forming a linear hydrogen-bonded 2:1 unit of the acid and the base. In (I), (II) and (III), inter-molecular C-H⋯O inter-actions are observed. The 2:1 units of (I) and (II) are linked via C-H⋯O hydrogen bonds, forming tape structures. In (III), the C-H⋯O hydrogen bonds, except for those formed in the units, link the two crystallographically independent 2:1 units. In (IV), no C-H⋯O inter-actions are observed, but π-π and C-H⋯π inter-actions link the units into a column structure.

  6. Microarray-based analysis and clinical validation identify ubiquitin-conjugating enzyme E2E1 (UBE2E1 as a prognostic factor in acute myeloid leukemia

    Directory of Open Access Journals (Sweden)

    Hongmei Luo

    2016-11-01

    Full Text Available Abstract Background Previous research suggested that single gene expression might be correlated with acute myeloid leukemia (AML survival. Therefore, we conducted a systematical analysis for AML prognostic gene expressions. Methods We performed a microarray-based analysis for correlations between gene expression and adult AML overall survival (OS using datasets GSE12417 and GSE8970. Positive findings were validated in an independent cohort of 50 newly diagnosed, non-acute promyelocytic leukemia (APL AML patients by quantitative RT-PCR and survival analysis. Results Microarray-based analysis suggested that expression of eight genes was each associated with 1-year and 3-year AML OS in both GSE12417 and GSE8970 datasets (p < 0.05. Next, we validated our findings in an independent cohort of AML samples collected in our hospital. We found that ubiquitin-conjugating enzyme E2E1 (UBE2E1 expression was adversely correlated with AML survival (p = 0.04. Multivariable analysis showed that UBE2E1 high patients had a significant shorter OS and shorter progression-free survival after adjusting other known prognostic factors (p = 0.03. At last, we found that UBE2E1 expression was negatively correlated with patients’ response to induction chemotherapy (p < 0.05. Conclusions In summary, we demonstrated that UBE2E1 expression was a novel prognostic factor in adult, non-APL AML patients.

  7. Reduction of benzene metabolism and toxicity in mice that lack CYP2E1 expression.

    Science.gov (United States)

    Valentine, J L; Lee, S S; Seaton, M J; Asgharian, B; Farris, G; Corton, J C; Gonzalez, F J; Medinsky, M A

    1996-11-01

    Transgenic CYP2E1 knockout mice (cyp2e1-/-) were used to investigate the involvement of CYP2E1 in the in vivo metabolism of benzene and in the development of benzene-induced toxicity. After benzene exposure, absence of CYP2E1 protein was confirmed by Western blot analysis of mouse liver samples. For the metabolism studies, male cyp2e1-/- and wild-type control mice were exposed to 200 ppm benzene, along with a radiolabeled tracer dose of [14C]benzene (1.0 Ci/mol) by nose-only inhalation for 6 hr. Total urinary radioactivity and all radiolabeled individual metabolites were reduced in urine of cyp2e1-/- mice compared to wild-type controls during the 48-hr period after benzene exposure. In addition, a significantly greater percentage of total urinary radioactivity could be accounted for as phenylsulfate conjugates in cyp2e1-/- mice compared to wild-type mice, indicating the importance of CYP2E1 in oxidation of phenol following benzene exposure in normal mice. For the toxicity studies, male cyp2e1-/-, wild-type, and B6C3F1 mice were exposed by whole-body inhalation to 0 ppm (control) or 200 ppm benzene, 6 hr/day for 5 days. On Day 5, blood, bone marrow, thymus, and spleen were removed for evaluation of micronuclei frequencies and tissue cellularities. No benzene-induced cytotoxicity or genotoxicity was observed in cyp2e1-/- mice. In contrast, benzene exposure resulted in severe genotoxicity and cytotoxicity in both wild-type and B6C3F1 mice. These studies conclusively demonstrate that CYP2E1 is the major determinant of in vivo benzene metabolism and benzene-induced myelotoxicity in mice.

  8. Characteristics and Soft-Demodulation of Galileo E1 Signal%Galileo E1信号特征及其软件解调

    Institute of Scientific and Technical Information of China (English)

    易翔

    2009-01-01

    Galileo E1民用信号采用新的BOC(1,1)扩频调制方式.针对E1信号进行信号建模,并对其信号特征进行了详细研究,最后设计了Galileo软件解调器中的信号处理模块.通过仿真验证表明,软件解调器成功实现了对Galileo E1信号的捕获、跟踪、解扩、解调全过程处理,克服了BOC解调的错锁问题.

  9. 26 CFR 1.669(e)-1A - Pro rata portion of taxes deemed distributed.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 8 2010-04-01 2010-04-01 false Pro rata portion of taxes deemed distributed. 1.669(e)-1A Section 1.669(e)-1A Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Treatment of Excess Distributions of Trusts Applicable to Taxable Years Beginning Before January...

  10. Expression of cytochrome P4502E1 gene in hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Xiao-Bo Man; Liang Tang; Xiu-Hua Qiu; Li-Qun Yang; Hui-Fang Cao; Meng-Chao Wu; Hong-Yang Wang

    2004-01-01

    AIM: To investigate cytpchrome P4502E1 ( CYP2E1) gene expression in occurrence and progression of hepatocellular carcinoma (HCC).METHODS: The human liver arrayed library was spotted onto the nylon membranes to make cDNA array. Hybridization of cDNA array was performed with labeled probes synthesized from RNA isolated from HCC and adjacent liver tissues.Sprague-Dawley rats were administrated diethylnitrosamine (DENA) to induce HCC. CYP2E1 expression was detected by the method of RT-PCR and Northern blot analysis.RESULTS: CYP2E1 was found by cDNA array hybridization to express differently between HCC and liver tissues. CYP2E1only expressed in liver, but did not express in HCC tissues and expressed lowly in cirrhotic tissues. In the progression of cirrhosis and HCC, the expression level of CYP2E1 was gradually decreased and hardly detected until the late stage of HCC.CONCLUSION: Using arrayed library to make cDNA arrays is an effective method to find differential expression genes.CYP2E1 is a unique gene expressing in liver but did not express in HCC. CYP2E1 expression descended along with the initiation and progression of HCC, which is noteworthy further investigations in its significance in the development of HCC.

  11. Morbidly Obese Patients Exhibit Increased CYP2E1-Mediated Oxidation of Acetaminophen

    NARCIS (Netherlands)

    A. van Rongen (Anne); P.A.J. Välitalo (Pyry A. J.); M.Y. Peeters (Mariska); D. Boerma (Djamila); F.W. Huisman (Fokko W.); B. van Ramshorst (Bert); E.H.P.A. van Dongen (Eric); J.N. van den Anker (John); C.A.J. Knibbe (Catherijne)

    2016-01-01

    textabstractIntroduction: Acetaminophen (paracetamol) is mainly metabolized via glucuronidation and sulphation, while the minor pathway through cytochrome P450 (CYP) 2E1 is held responsible for hepatotoxicity. In obese patients, CYP2E1 activity is reported to be induced, thereby potentially worsenin

  12. Morbidly Obese Patients Exhibit Increased CYP2E1-Mediated Oxidation of Acetaminophen.

    NARCIS (Netherlands)

    Rongen, van A.; Välitalo, P.A.J.; Peeters, M.Y.; Boerma, D.; Huisman, F.W.; Ramshorst, van B.; Dongen, van E.P.; Anker, van den J.N.; Knibbe, C.A.J.

    2016-01-01

    INTRODUCTION Acetaminophen (paracetamol) is mainly metabolized via glucuronidation and sulphation, while the minor pathway through cytochrome P450 (CYP) 2E1 is held responsible for hepatotoxicity. In obese patients, CYP2E1 activity is reported to be induced, thereby potentially worsening the safety

  13. CK2 phosphorylation inactivates DNA binding by the papillomavirus E1 and E2 proteins.

    Science.gov (United States)

    Schuck, Stephen; Ruse, Cristian; Stenlund, Arne

    2013-07-01

    Papillomaviruses have complex life cycles that are understood only superficially. Although it is well established that the viral E1 and E2 proteins play key roles in controlling viral transcription and DNA replication, how these factors are regulated is not well understood. Here, we demonstrate that phosphorylation by the protein kinase CK2 controls the biochemical activities of the bovine papillomavirus E1 and E2 proteins by modifying their DNA binding activity. Phosphorylation at multiple sites in the N-terminal domain in E1 results in the loss of sequence-specific DNA binding activity, a feature that is also conserved in human papillomavirus (HPV) E1 proteins. The bovine papillomavirus (BPV) E2 protein, when phosphorylated by CK2 on two specific sites in the hinge, also loses its site-specific DNA binding activity. Mutation of these sites in E2 results in greatly increased levels of latent viral DNA replication, indicating that CK2 phosphorylation of E2 is a negative regulator of viral DNA replication during latent viral replication. In contrast, mutation of the N-terminal phosphorylation sites in E1 has no effect on latent viral DNA replication. We propose that the phosphorylation of the N terminus of E1 plays a role only in vegetative viral DNA replication, and consistent with such a role, caspase 3 cleavage of E1, which has been shown to be necessary for vegetative viral DNA replication, restores the DNA binding activity to phosphorylated E1.

  14. 26 CFR 1.665(e)-1A - Preceding taxable year.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 8 2010-04-01 2010-04-01 false Preceding taxable year. 1.665(e)-1A Section 1... (CONTINUED) INCOME TAXES Treatment of Excess Distributions of Trusts Applicable to Taxable Years Beginning on Or After January 1, 1969 § 1.665(e)-1A Preceding taxable year. (a) Definition—(1) Domestic trusts—(i...

  15. New Protein Vector ApE1 for Targeted Delivery of Anticancer Drugs

    Directory of Open Access Journals (Sweden)

    N. V. Pozdniakova

    2012-01-01

    Full Text Available A new chimeric gene ApE1 encoding the receptor-binding domain of the human alpha-fetoprotein fused to a sequence of 22 glutamic acid residues was constructed. A new bacterial producer strain E. coli SHExT7 ApE1 was selected for ApE1 production in a soluble state. A simplified method was developed to purify ApE1 from bacterial biomass. It was shown that the new vector protein selectively interacts with AFP receptors on the tumor cell surface and can be efficiently accumulated in tumor cells. In addition, ApE1 was shown to be stable in storage and during its chemical modification. An increased number of carboxyl groups in the molecule allows the production of cytotoxic compound conjugates with higher drug-loading capacity and enhanced tumor targeting potential.

  16. Comparison of osteogenic potentials of human rat BMP4 and BMP6 gene therapy using [E1-] and [E1-,E2b-] adenoviral vectors

    Directory of Open Access Journals (Sweden)

    Hongwei Li, Jin Zhong Li, Debra D. Pittman, Andy Amalfitano, Gerald R. Hankins, Gregory A. Helm

    2006-01-01

    Full Text Available Osteogenic potentials of some recombinant human bone morphogenetic protein (BMP first-generation adenoviral vectors (ADhBMPs are significantly limited in immunocompetent animals. It is unclear what role expression of viral proteins and foreign proteins transduced by adenoviral vectors play in the host immune response and in ectopic bone formation. In this study two sets of experiments were designed and performed. First, rat BMP6 cDNA were amplified, sequenced, and recombined in first-generation adenoviral vector (ADrBMP6. A comparison of human and rat BMP6 adenoviral vectors demonstrated identical osteogenic activities in both immunodeficient and immunocompetent rats. Second, the activities of recombinant human BMP6 in E1- (ADhBMP6 and [E1-,E2b-] ( [E1-,E2b-]ADGFP&hBMP6, and [E1-,E2b-]ADhBMP6 adenoviral vectors were compared in both in vitro and in vivo models. Similar activities of these two generations of BMP adenoviral vectors were found in all models. These results indicate that the amount of viral gene expression and the source of the BMP cDNA are not major factors in the interruption of osteogenic potentials of recombinant BMP6 adenoviral vectors in immunocompetent animals.

  17. A design for clock data recovery circuit of E1%一种E1时钟数据恢复电路的设计

    Institute of Scientific and Technical Information of China (English)

    李鑫; 黄海生; 张斌; 惠楠

    2012-01-01

    针对E1数据的时钟数据恢复问题,设计一种基于小数分频且有环路滤波功能的数控振荡器(DigitallyControlled Oscillator,DCO),给出一种新的全数字锁相环(All Digital Phase-Locked Loop,ADPLL)实现方案,将数字环路滤波器(Digital Loop Filter,DLF)和DCO集成到一个模块,从而实现一种E1时钟数据恢复(Clock Data Re-covery,CDR)电路。经过对比可知,新方案比传统ADPLL实现方案的电路集成度更高。理论分析显示,新方案电路性能可靠。%For the problem of E1 clock data recovery, a kind of Digitally Controlled Oscillator (DCO), which has loop filtering function based on fractional frequency dividing, is presented. Then it gives a new kind of All Digital Phase-Locked Loop(ADPLL) implemention scheme. On the basis of this scheme, a kind of E1 Clock Data Reeovery(CDR) circuit is implemented. Com- pared with traditional ADPLL implemention scheme, the scheme presented integrates Digital Loop Filter (DLF) and DCO together, thus the integration of the circuit becomes higher. The mathematical analysis to the circuit performance proves that the design can reliably recover the data and clock from E1 data received.

  18. Long-Term Prostaglandin E1 Infusion for Newborns with Critical Congenital Heart Disease.

    Science.gov (United States)

    Aykanat, Alper; Yavuz, Taner; Özalkaya, Elif; Topçuoğlu, Sevilay; Ovalı, Fahri; Karatekin, Güner

    2016-01-01

    Prostaglandin E1 is crucial for keeping the patent ductus arteriosus in critical congenital heart disease for the survival and palliation of particularly prematurely born babies until a cardiosurgical intervention is available. In this study, the side effects of prostaglandin E1 in newborns with critical congenital heart disease and clinical outcomes were evaluated. Thirty-five newborns diagnosed with critical congenital heart disease were treated with prostaglandin E1 between January 2012 and September 2014 at our hospital. Patient charts were examined for prostaglandin E1 side effects (metabolic, gastric outlet obstruction, apnea), clinical status, and prognosis. Acquired data were analyzed in the SPSS 20.0 program. Patients with birth weight under 2500 g needed more days of prostaglandin E1 infusion than ones with birthweight over 2500 g (P = 0.016). The ratio of patients with birth weight under 2500 g who received prostaglandin E1 longer than 7 days was higher than the patients with birth weight over 2500 g (P = 0.02). Eighteen side effects were encountered in 11 of 35 patients (31%). Of these side effects, 1 patient had 4, 4 patients had 2, and 6 patients had only 1 side effect. Discontinuation of the therapy was never needed. Prostaglandin E1 is an accepted therapy modality for survival and outcome in critical congenital heart disease in particularly low-birth-weight babies until a surgical intervention is available. Side effects are not less encountered but are almost always manageable, and discontinuation is not needed.

  19. Candida albicans modulates host defense by biosynthesizing the pro-resolving mediator resolvin E1.

    Directory of Open Access Journals (Sweden)

    Eric J Haas-Stapleton

    Full Text Available Candida albicans is an opportunistic fungal pathogen of humans that resides commensally on epithelial surfaces, but can cause inflammation when pathogenic. Resolvins are a class of anti-inflammatory lipids derived from omega-3 polyunsaturated fatty acids (PUFA that attenuate neutrophil migration during the resolution phase of inflammation. In this report we demonstrate that C. albicans biosynthesizes resolvins that are chemically identical to those produced by human cells. In contrast to the trans-cellular biosynthesis of human Resolvin E1 (RvE1, RvE1 biosynthesis in C. albicans occurs in the absence of other cellular partners. C. albicans biosynthesis of RvE1 is sensitive to lipoxygenase and cytochrome P450 monoxygenase inhibitors. We show that 10nM RvE1 reduces neutrophil chemotaxis in response to IL-8; 1nM RvE1 enhanced phagocytosis of Candida by human neutrophils, as well as intracellular ROS generation and killing, while having no direct affect on neutrophil motility. In a mouse model of systemic candidiasis, RvE1 stimulated clearance of the fungus from circulating blood. These results reveal an inter-species chemical signaling system that modulates host immune functions and may play a role in balancing host carriage of commensal and pathogenic C. albicans.

  20. E1DS: catalytic site prediction based on 1D signatures of concurrent conservation.

    Science.gov (United States)

    Chien, Ting-Ying; Chang, Darby Tien-Hao; Chen, Chien-Yu; Weng, Yi-Zhong; Hsu, Chen-Ming

    2008-07-01

    Large-scale automatic annotation of protein sequences remains challenging in postgenomics era. E1DS is designed for annotating enzyme sequences based on a repository of 1D signatures. The employed sequence signatures are derived using a novel pattern mining approach that discovers long motifs consisted of several sequential blocks (conserved segments). Each of the sequential blocks is considerably conserved among the protein members of an EC group. Moreover, a signature includes at least three sequential blocks that are concurrently conserved, i.e. frequently observed together in sequences. In other words, a sequence signature is consisted of residues from multiple regions of the protein sequence, which echoes the observation that an enzyme catalytic site is usually constituted of residues that are largely separated in the sequence. E1DS currently contains 5421 sequence signatures that in total cover 932 4-digital EC numbers. E1DS is evaluated based on a collection of enzymes with catalytic sites annotated in Catalytic Site Atlas. When compared to the famous pattern database PROSITE, predictions based on E1DS signatures are considered more sensitive in identifying catalytic sites and the involved residues. E1DS is available at http://e1ds.ee.ncku.edu.tw/ and a mirror site can be found at http://e1ds.csbb.ntu.edu.tw/.

  1. Enterasys Networks delivers 10-Gigabit ethernet for the enterprise with new matrix E1 switching family

    CERN Multimedia

    2001-01-01

    Enterasys Networks Inc., today announced its new Matrix E1 family of 10-Gigabit and Gigabit Ethernet switches. The Matrix E1 Optical Access Switch (OAS) enables organizations to deliver applications at 10-Gb speeds across a single fibre optic pair. Jacques Altaber, deputy leader of IT at CERN said "High-bandwith solutions are essential to leveraging more computing power, so 10-Gb Ethernet is the next logical step for us...The Matrix E1 allows us to provide the networking support that our scientists need and gives us a certain future for bandwidth and computing expansion".

  2. Inactivation of cyclin E1 inhibits chemically induced hepatocarcinogenesis in mice

    OpenAIRE

    Moro, Nives

    2011-01-01

    E-type cyclins (CcnE) control the transition of quiescent cells into the cell cycle. Two E-type cyclins, CcnE1 and CcnE2 have been described. A variety of human cancers, including hepatocellular carcinoma (HCC), overexpress CcnE and this is frequently associated with reduced patient survival. The aim of the present study was to dissect the role of CcnE1 and CcnE2 for hepatocarcinogenesis induced by the carcinogen diethylnitrosamine (DEN) using CcnE1 and CcnE2 knockout mice. The central questi...

  3. Acetato(N-[(E-1-(6-methyl-2-pyridylmethylidene]-2-{2-[(E-1-(6-methyl-2-pyridylmethylideneamino]phenethyl}anilinenickel(II perchlorate

    Directory of Open Access Journals (Sweden)

    Hong-In Lee

    2010-10-01

    Full Text Available In the title complex, [Ni(CH3COO(C28H26N4]ClO4, the NiII atom is coordinated by two imine N atoms and two pyridine N atoms of the N-[(E-1-(6-methyl-2-pyridylmethylidene]-2-(2-[(E-1-(6-methyl-2-pyridylmethylideneamino]phenethylaniline donor ligand and two O atoms of the acetate ion in a distorted octahedral coordination. The average Ni—N and Ni—O bond lengths are 2.131 (13 and 2.098 (11 Å, respectively. An intramolecular N—H...O interaction occurs. Relatively weak intermolecular C—H...O interactions between the ligands and the ClO4− ions result in a chain extending along the b axis.

  4. E1, M1, E2 transition energies and probabilities of W$^{54+}$ ions

    CERN Document Server

    Ding, Xiao-bin; Liu, Jia-xin; Koike, Fumihiro; Murakami, Izumi; Kato, Daiji; Sakaue, Hiroyuki A; Nakamura, Nobuyuki; Dong, Chen-zhong

    2016-01-01

    A comprehensive theoretical study of the E1, M1, E2 transitions of Ca-like tungsten ions is presented. Using multi-configuration Dirac-Fock (MCDF) method with a restricted active space treatment, the wavelengths and probabilities of the M1 and E2 transitions between the multiplets of the ground state configuration ([Ne]3s$^{2}$3p$^{6}$3d$^{2}$) and of the E1 transitions between [Ne]3s$^{2}$3p$^{5}$3d$^{3}$ and [Ne]3s$^{2}$3p$^{6}$3d$^{2}$ have been calculated. The results are in reasonable agreement with available experimental data. The present E1 and M1 calculations are compared with previous theoretical values. For E2 transitions, the importance of electron correlation from 3s and 3p orbitals is pointed out. Several strong E1 transitions are predicted, which have potential advantage for plasma diagnostics.

  5. 26 CFR 1.1397E-1 - Qualified zone academy bonds.

    Science.gov (United States)

    2010-04-01

    ... TAX (CONTINUED) INCOME TAXES Empowerment Zone Employment Credit § 1.1397E-1 Qualified zone academy...), (iii), (iv) or (v). Services of employees of the eligible local education agency do not constitute...

  6. 78 FR 15597 - Special Conditions: GE Aviation CT7-2E1 Turboshaft Engine Model

    Science.gov (United States)

    2013-03-12

    ... conditions are issued for the General Electric Aviation (GE) CT7-2E1 engine model. This engine model will... ensure the test will run for a 2.5 minute duration with no interruption. Type Certification Basis Under...

  7. Effects of E2 and E1-E2 Interference on Coulomb Dissociation of 19C

    Institute of Scientific and Technical Information of China (English)

    Rajesh Kharab; Pardeep Singh; Ravinder Kumar

    2007-01-01

    We investigate the effects of higher order multipole transitions, in particular electric quadrupole (E2) and E1-E2interference, on the Coulomb dissociation of 19 C within the framework of the first order eikonal approximation.The sensitivity of the total Coulomb breakup cross section and the longitudinal momentum distribution of the core fragment to these effects are checked. The breakup occurs predominately through the dipole transition and the contribution of E2 transition to the total cross section is found to be within the range from 1 to 3% of that of E1. It is further observed that the E1-E2 interference term contributes nothing to the integrated cross section.On the other hand, the longitudinal momentum distribution is observed to be insensitive to the E2 transition while the E1-E2 interference introduces a small asymmetry in its shape.

  8. The C-terminal region of E1A: a molecular tool for cellular cartography.

    Science.gov (United States)

    Yousef, Ahmed F; Fonseca, Gregory J; Cohen, Michael J; Mymryk, Joe S

    2012-04-01

    The adenovirus E1A proteins function via protein-protein interactions. By making many connections with the cellular protein network, individual modules of this virally encoded hub reprogram numerous aspects of cell function and behavior. Although many of these interactions have been thoroughly studied, those mediated by the C-terminal region of E1A are less well understood. This review focuses on how this region of E1A affects cell cycle progression, apoptosis, senescence, transformation, and conversion of cells to an epithelial state through interactions with CTBP1/2, DYRK1A/B, FOXK1/2, and importin-α. Furthermore, novel potential pathways that the C-terminus of E1A influences through these connections with the cellular interaction network are discussed.

  9. CYP2E1-catalyzed oxidation contributes to the sperm toxicity of 1-bromopropane in mice.

    Science.gov (United States)

    Garner, C Edwin; Sloan, C; Sumner, S C J; Burgess, J; Davis, J; Etheridge, A; Parham, A; Ghanayem, B I

    2007-03-01

    1-bromopropane (1-BrP) induces dose- and time-dependent reproductive organ toxicity and reduced sperm motility in rodents. The contribution of cytochrome P4502E1 (CYP2E1) to both 1-BrP metabolism and the induction of male reproductive toxicity was investigated using wild-type (WT) and Cyp2e1-/- mice. In gas uptake inhalation studies, the elimination half-life of [1,2,3-(13)C]-1-BrP was longer in Cyp2e1-/- mice relative to WT (3.2 vs. 1.3 h). Urinary metabolites were identified by 13C nuclear magnetic resonance. The mercapturic acid of 1-bromo-2-hydroxypropane (2OHBrP) was the major urinary metabolite in WT mice, and products of conjugation of 1-BrP with glutathione (GSH) were insignificant. The ratio of GSH conjugation to 2-hydroxylation increased 5-fold in Cyp2e1-/- mice relative to WT. After 1-BrP exposure, hepatic GSH was decreased by 76% in WT mice vs. 47% in Cyp2e1-/- mice. Despite a 170% increase in 1-BrP exposure in Cyp2e1-/- vs. WT mice, sperm motility in exposed Cyp2e1-/- mice did not change relative to unexposed matched controls. This suggests that metabolites produced through CYP2E1-mediated oxidation may be responsible for 1-BrP-induced sperm toxicity. Both 1-BrP and 2OHBrP inhibited the motility of sperm obtained from WT mice in vitro. However, only 2OHBrP reduced the motility of sperm obtained from Cyp2e1-/- mice in vitro, suggesting that conversion of parent compound to 2OHBrP within the spermatozoa may contribute, at least in part, to reduced motility. Overall, these data suggest that metabolism of 1-BrP is mediated in part by CYP2E1, and activation of 1BrP via this enzyme may contribute to the male reproductive toxicity of this chemical.

  10. Desmodesmus sp. 3Dp86E-1-a novel symbiotic chlorophyte capable of growth on pure CO2.

    Science.gov (United States)

    Solovchenko, Alexei; Gorelova, Olga; Selyakh, Irina; Semenova, Larisa; Chivkunova, Olga; Baulina, Olga; Lobakova, Elena

    2014-10-01

    A novel chlorophyte Desmodesmus sp. 3Dp86E-1 isolated from a White Sea hydroid Dynamena pumila was cultivated at CO2 levels from atmospheric (the 'low-CO2' conditions) to pure carbon dioxide (the 5, 20, and 100 % CO2 conditions) under high (480 μE/(m(2) s) PAR) light. After 7 days of cultivation, the '100 % CO2' (but not 5 or 20 % CO2) cells possessed ca. four times higher chlorophyll content per dry weight (DW) unit than the low-CO2 culture. The rate of CO2 fixation under 100 % CO2 comprised ca. 1.5 L/day per L culture volume. After a lag period which depended on the CO2 level, biomass accumulation and volumetric fatty acid (FA) content of the Desmodesmus sp. 3Dp86E-1 bubbled with CO2-enriched gas mixtures increased and was comparable to that of the culture continuously bubbled with air. Under the low-to-moderate CO2 conditions, the FA percentage of the algal cells increased (to 40 % DW) whereas under high-CO2 conditions, FA percentage did not exceed 15 % DW. A strong increase in oleate (18:1) proportion of total FA at the expense of linolenate (18:3) was recorded in the '100 % CO2' cells. Electron microscopy and pulse-amplitude-modulated chlorophyll fluorescence investigation revealed no damage to or significant downregulation of the photosynthetic apparatus in '100 % CO2' cells grown at the high-PAR irradiance. Possible mechanisms of high-CO2 tolerance of Desmodesmus sp. 3Dp86E-1 are discussed in view of its symbiotic origin and possible application for CO2 biomitigation.

  11. Role of CYP2E1 gene polymorphisms association with hepatitis risk in Northeast India

    Institute of Scientific and Technical Information of China (English)

    Manab; Deka; Moumita; Bose; Bharati; Baruah; Purabi; Deka; Bose; Subhash; Medhi; Sujoy; Bose; Anjan; Saikia; Premashish; Kar

    2010-01-01

    AIM:To investigate hepatitis virus, genetic and environmental factors, and their interactions in predisposing patients to liver diseases in Northeast India. METHODS:A total of 104 jaundice patients and 124 community controls were included. Serological analysis was performed by routine enzyme-linked immunosorbent assay, and nucleic acid testing for hepatitis viruses was done by polymerase chain reaction (PCR), followed by PCR direct sequencing for viral genotyping. Cytochrome P450 2E1 (CYP2E1) polymorphism w...

  12. Cooperative effects for CYP2E1 differ between styrene and its metabolites

    OpenAIRE

    2013-01-01

    Cooperative interactions are frequently observed in the metabolism of drugs and pollutants by cytochrome P450s; nevertheless, the molecular determinants for cooperativity remain elusive. Previously, we demonstrated that steady-state styrene metabolism by CYP2E1 exhibits positive cooperativity.We hypothesized that styrene metabolites have lower affinity than styrene toward CYP2E1 and limited ability to induce cooperative effects during metabolism. To test the hypothesis, we determined the pote...

  13. Phenotypic diversity of toxigenic Vibrio cholerae O1 E1 Tor strains identified in China

    Institute of Scientific and Technical Information of China (English)

    赵璇

    2014-01-01

    Objective To understand the phenotypic diversity of toxigenic Vibrio cholerae O1 E1 Tor strains isolated from different provinces in China during the last 50 years.Methods Traditional biotyping testings including susceptibility to polymyxin B,sensitivity to groupⅣphage,Voges-Proskauer test and haemolysis of sheep erythrocytes were conducted.Results Data from Biotype-specific phenotype analysis revealed that only 133 isolates carried the typical E1 Tor phenotypes while the other 251

  14. Evolution of the ubiquitin-activating enzyme Uba1 (E1)

    Science.gov (United States)

    Allan, Douglas C.; Phillips, J. C.

    2017-10-01

    Ubiquitin tags diseased proteins and initiates an enzyme conjugation cascade, which has three stages. The first-stage enzyme Uba1 (E1) has evolved only modestly from slime mold to humans, and is > 14 times larger than Ub. Here we use critical point thermodynamic scaling theory to connect Uba1 (E1) evolution from yeast and slime mold to fruit flies and humans to subtle changes in its amino acid sequences.

  15. The effect of prostaglandin E_1 on recovery of early renal graft functions after transplantation

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Objective To investigate the effect of prostaglandin E1 (PGE1) on recovery of early renal graft functions after transplantation. Methods One hundred and seven patients after renal transplantation were allocated in the treated group, and treated by conventional treatment with injection of 10 μg prostaglandin E1 additionally twice a day for 14 days. And eighty-eight patients who received conventional treatment alone after renal transplantation at the corresponding period were allocated in the control group. I...

  16. The Frequency of Cytochrome P450 2E1 Polymorphisms in Black South Africans

    Directory of Open Access Journals (Sweden)

    Paul K. Chelule

    2006-01-01

    Full Text Available Polymorphisms in the promoter region of the Cytochrome P4502E1 (CYP2E1 gene reportedly modify the metabolic activity of CYP2E1 enzyme, and have been associated with increased susceptibility to squamous cell carcinoma (SCC of the oesophagus in high prevalence areas such as China. To assess the frequency of these polymorphisms in Black South Africans, a population with a high incidence of oesophageal SCC, this study examined genomic DNA from 331 subjects for restriction fragment length polymorphisms in the CYP2E1 (RsaI and PstI digestion. The frequency of the CYP2E1 c1/c1 and c1/c3 genotypes was 95% and 5% respectively. The frequency of the CYP2E1 allele distribution was found to be markedly different between Chinese and South African populations; hence it is important to place racial differences into consideration when proposing allelic variants as genetic markers for cancer.

  17. In vitro characterization of the NAD+ synthetase NadE1 from Herbaspirillum seropedicae.

    Science.gov (United States)

    Laskoski, Kerly; Santos, Adrian R S; Bonatto, Ana C; Pedrosa, Fábio O; Souza, Emanuel M; Huergo, Luciano F

    2016-05-01

    Nicotinamide adenine dinucleotide synthetase enzyme (NadE) catalyzes the amination of nicotinic acid adenine dinucleotide (NaAD) to form NAD(+). This reaction represents the last step in the majority of the NAD(+) biosynthetic routes described to date. NadE enzymes typically use either glutamine or ammonium as amine nitrogen donor, and the reaction is energetically driven by ATP hydrolysis. Given the key role of NAD(+) in bacterial metabolism, NadE has attracted considerable interest as a potential target for the development of novel antibiotics. The plant-associative nitrogen-fixing bacteria Herbaspirillum seropedicae encodes two putative NadE, namely nadE1 and nadE2. The nadE1 gene is linked to glnB encoding the signal transduction protein GlnB. Here we report the purification and in vitro characterization of H. seropedicae NadE1. Gel filtration chromatography analysis suggests that NadE1 is an octamer. The NadE1 activity was assayed in vitro, and the Michaelis-Menten constants for substrates NaAD, ATP, glutamine and ammonium were determined. Enzyme kinetic and in vitro substrate competition assays indicate that H. seropedicae NadE1 uses glutamine as a preferential nitrogen donor.

  18. Structural analysis and evolution of specificity of the SUMO UFD E1-E2 interactions

    Science.gov (United States)

    Liu, Bing; Lois, L. Maria; Reverter, David

    2017-01-01

    SUMO belongs to the ubiquitin-like family (UbL) of protein modifiers. SUMO is conserved among eukaryotes and is essential for the regulation of processes such as DNA damage repair, transcription, DNA replication and mitosis. UbL modification of proteins occurs via a specific enzymatic cascade formed by the crosstalk between the E1-activating enzyme, the E2-conjugating enzyme and the E3-ligase. An essential discrimination step in all UbL modifiers corresponds to the interaction between E1 and E2 enzymes, which is mediated by the recruitment of the E2 to the UFD domain (Ubiquitin-Fold Domain) of the E1 enzyme. To gain insights in the properties of this interface, we have compared the structures of the complexes between E1 UFD domain and E2 in human and yeast, revealing two alternative UFD platforms that interact with a conserved E2. Comparative sequence analysis of the E1 UFD domain indicates that the E2 binding region has been conserved across phylogenetic closely related species, in which higher sequence conservation can be found in the E2 binding region than in the entire UFD domain. These distinctive strategies for E1-E2 interactions through the UFD domain might be the consequence of a high selective pressure to ensure specificity of each modifier conjugation system. PMID:28165030

  19. Autophagy Protects against CYP2E1/Chronic Ethanol-Induced Hepatotoxicity

    Directory of Open Access Journals (Sweden)

    Yongke Lu

    2015-10-01

    Full Text Available Autophagy is an intracellular pathway by which lysosomes degrade and recycle long-lived proteins and cellular organelles. The effects of ethanol on autophagy are complex but recent studies have shown that autophagy serves a protective function against ethanol-induced liver injury. Autophagy was found to also be protective against CYP2E1-dependent toxicity in vitro in HepG2 cells which express CYP2E1 and in vivo in an acute alcohol/CYPE1-dependent liver injury model. The goal of the current report was to extend the previous in vitro and acute in vivo experiments to a chronic ethanol model to evaluate whether autophagy is also protective against CYP2E1-dependent liver injury in a chronic ethanol-fed mouse model. Wild type (WT, CYP2E1 knockout (KO or CYP2E1 humanized transgenic knockin (KI, mice were fed an ethanol liquid diet or control dextrose diet for four weeks. In the last week, some mice received either saline or 3-methyladenine (3-MA, an inhibitor of autophagy, or rapamycin, which stimulates autophagy. Inhibition of autophagy by 3-MA potentiated the ethanol-induced increases in serum transaminase and triglyceride levels in the WT and KI mice but not KO mice, while rapamycin prevented the ethanol liver injury. Treatment with 3-MA enhanced the ethanol-induced fat accumulation in WT mice and caused necrosis in the KI mice; little or no effect was found in the ethanol-fed KO mice or any of the dextrose-fed mice. 3-MA treatment further lowered the ethanol-decrease in hepatic GSH levels and further increased formation of TBARS in WT and KI mice, whereas rapamycin blunted these effects of ethanol. Neither 3-MA nor rapamycin treatment affected CYP2E1 catalytic activity or content or the induction CYP2E1 by ethanol. The 3-MA treatment decreased levels of Beclin-1 and Atg 7 but increased levels of p62 in the ethanol-fed WT and KI mice whereas rapamycin had the opposite effects, validating inhibition and stimulation of autophagy, respectively. These

  20. Cytochrome p450 2E1 polymorphisms and the risk of gastric cardia cancer

    Institute of Scientific and Technical Information of China (English)

    Lin Cai; Zong-Li Zheng; Zuo-Feng Zhang

    2005-01-01

    AIM: Genetic polymorphisms of drug-metabolizing enzymes have recently been shown to affect susceptibility to chemical carcinogenesis. Cytochrome P450 2E1 (CYP2E1)enzyme catalyzes the metabolism of many procarcinogens,such as N-nitrosamines and related compounds. The gene coding for this enzyme is polymorphic and thus may play a role in gastric cardia cancer (GCC) etiology. In this hospital-based case-control study, we evaluate the relationship between genetic polymorphisms of CYP2E1and the risk of GCC.METHODS: The study subjects comprised 159 histologically confirmed GCC cases identified via hospital cancer registry and surgical records at five hospitals in Fuzhou, Fujian Province, China, between April and November 2001.Controls were 192 patients admitted to the same hospitals for nonmalignant conditions. The genotypes of CYP2E1were detected by a PCR-based RFLP assay. The odds ratios were estimated by logistic regression analyses and were adjusted for potential confounding factors.RESULTS: The distribution of three genotypes of CYP2E1in GCC cases and controls was significantly different (X2 = 16.04, P<0.01). The frequency of the CYP2E1 (c1/c1)genotype in GCC cases and controls was 60.4% and 40.1%,respectively. The CYP2E1 (c1/c1) genotype was associated with an increased risk for GCC (the adjusted (OR) was 2.37, 95% confidence interval (CI): 1.52-3.70). Subjects who carried the CYP2E 1 (c1/c1) genotype and were habitual smokers were at a significantly higher risk of developing GCC (OR = 4.68, 95%CI: 2.19-10.04) compared with those who had the CYP2E1 (c1/c2or c2/c2') genotype and did not smoke.CONCLUSION: These results suggest that the CYP2E 1genotype may influence individual susceptibility to development of GCC, and that the risk increases significantly in smokers.

  1. A shock-tube measurement of the SiO/E 1 Sigma + - X 1 Sigma +/ transition moment

    Science.gov (United States)

    Park, C.

    1978-01-01

    The sum of the squares of the electronic transition moments for the (E 1 Sigma +) - (X 1 Sigma +) band system of SiO has been determined from absorption measurements conducted in the reflected-shock region of a shock tube. The test gas produced by shock-heating a mixture of SiCl4, N2O, and Ar, and the spectra were recorded photographically in the 150-230-nm wavelength range. The values of the sum of the squares were determined by comparing the measured absorption spectra with those produced by a line-by-line synthetic spectrum calculation. The value so deduced at an r-centroid value of 3.0 bohr was 0.86 + or - 0.10 atomic unit.

  2. Expression of Estrogen Sulfotransferase 1E1 and Steroid Sulfatase in Breast Cancer: A Immunohistochemical Study

    Directory of Open Access Journals (Sweden)

    D. Poisson Paré

    2009-01-01

    Full Text Available It is known that the steroid sulfatase (STS and the estrogen sulfotransferase (EST1E1 are commonly expressed in human breast carcinomas. STS and EST1E1 combined action could maintain the equilibrium between sulfated (inactive and unconjugated (active estrogens, which might have effects on development of hormone dependent breast cancer. We studied the expression of the STS and EST1E1 in 88 breast carcinomas and 57 adjacent non-malignant tissues by immunohistochemistry. The results were correlated with the tumor expression of estrogen receptor α (ER-α and β (ER-β, progesterone receptor A (PR-A and B (PR-B and the proliferation marker CDC47, the tumoral type and stage and the age at surgery. STS expression was higher in carcinoma specimens than in adjacent normal tissues, although not to a significant level (p = 0.064 and it was positively associated with CDC47 expression (p 0.05. These observations support the hypothesis that STS is overexpressed in breast cancer and associated with a worse prognosis. EST1E1 was observed for the first time in the nuclei of epithelial and tumoral cells. Tumor expression of EST1E1 was positively correlated with ER-β (p 0.01 and PR-B (p 0.05, two steroid receptors already associated with an improve prognosis for breast cancer. Controlling the STS overexpression in carcinomas could be a way to inhibit cancer growth. The significance of the association between EST1E1 and ER-β or PR-B should be further studied since these two receptors are transcription activators and may regulate the expression of protective enzymes like EST1E1.

  3. Mutation in E1, the ubiquitin activating enzyme, reduces Drosophila lifespan and results in motor impairment.

    Science.gov (United States)

    Liu, Hsiu-Yu; Pfleger, Cathie M

    2013-01-01

    Neurodegenerative diseases cause tremendous suffering for those afflicted and their families. Many of these diseases involve accumulation of mis-folded or aggregated proteins thought to play a causal role in disease pathology. Ubiquitinated proteins are often found in these protein aggregates, and the aggregates themselves have been shown to inhibit the activity of the proteasome. These and other alterations in the Ubiquitin Pathway observed in neurodegenerative diseases have led to the question of whether impairment of the Ubiquitin Pathway on its own can increase mortality or if ongoing neurodegeneration alters Ubiquitin Pathway function as a side-effect. To address the role of the Ubiquitin Pathway in vivo, we studied loss-of-function mutations in the Drosophila Ubiquitin Activating Enzyme, Uba1 or E1, the most upstream enzyme in the Ubiquitin Pathway. Loss of only one functional copy of E1 caused a significant reduction in adult lifespan. Rare homozygous hypomorphic E1 mutants reached adulthood. These mutants exhibited further reduced lifespan and showed inappropriate Ras activation in the brain. Removing just one functional copy of Ras restored the lifespan of heterozygous E1 mutants to that of wild-type flies and increased the survival of homozygous E1 mutants. E1 homozygous mutants also showed severe motor impairment. Our findings suggest that processes that impair the Ubiquitin Pathway are sufficient to cause early mortality. Reduced lifespan and motor impairment are seen in the human disease X-linked Infantile Spinal Muscular Atrophy, which is associated with mutation in human E1 warranting further analysis of these mutants as a potential animal model for study of this disease.

  4. Cytochrome P450-2E1 promotes fast food-mediated hepatic fibrosis

    Science.gov (United States)

    Abdelmegeed, Mohamed A.; Choi, Youngshim; Godlewski, Grzegorz; Ha, Seung-Kwon; Banerjee, Atrayee; Jang, Sehwan; Song, Byoung-Joon

    2017-01-01

    Cytochrome P450-2E1 (CYP2E1) increases oxidative stress. High hepatic cholesterol causes non-alcoholic steatohepatitis (NASH) and fibrosis. Thus, we aimed to study the role of CYP2E1 in promoting liver fibrosis by high cholesterol-containing fast-food (FF). Male wild-type (WT) and Cyp2e1-null mice were fed standard chow or FF for 2, 12, and 24 weeks. Various parameters of liver fibrosis and potential mechanisms such as oxidative and endoplasmic reticulum (ER) stress, inflammation, and insulin resistance (IR) were studied. Indirect calorimetry was also used to determine metabolic parameters. Liver histology showed that only WT fed FF (WT-FF) developed NASH and fibrosis. Hepatic levels of fibrosis protein markers were significantly increased in WT-FF. The nitroxidative stress marker iNOS, but not CYP2E1, was significantly elevated only in FF-fed WT. Serum endotoxin, TLR-4 levels, and inflammatory markers were highest in WT-FF. FAS, PPAR-α, PPAR-γ, and CB1-R were markedly altered in WT-FF. Electron microscopy and immunoblot analyses showed significantly higher levels of ER stress in FF-fed WT. Indirect calorimetry showed that Cyp2e1-null-mice fed FF exhibited consistently higher total energy expenditure (TEE) than their corresponding WT. These results demonstrate that CYP2E1 is important in fast food-mediated liver fibrosis by promoting nitroxidative and ER stress, endotoxemia, inflammation, IR, and low TEE. PMID:28051126

  5. CYP2E1-dependent elevation of serum cholesterol, triglycerides, and hepatic bile acids by isoniazid.

    Science.gov (United States)

    Cheng, Jie; Krausz, Kristopher W; Li, Feng; Ma, Xiaochao; Gonzalez, Frank J

    2013-01-15

    Isoniazid is the first-line medication in the prevention and treatment of tuberculosis. Isoniazid is known to have a biphasic effect on the inhibition-induction of CYP2E1 and is also considered to be involved in isoniazid-induced hepatotoxicity. However, the full extent and mechanism of involvement of CYP2E1 in isoniazid-induced hepatotoxicity remain to be thoroughly investigated. In the current study, isoniazid was administered to wild-type and Cyp2e1-null mice to investigate the potential toxicity of isoniazid in vivo. The results revealed that isoniazid caused no hepatotoxicity in wild-type and Cyp2e1-null mice, but produced elevated serum cholesterol and triglycerides, and hepatic bile acids in wild-type mice, as well as decreased abundance of free fatty acids in wild-type mice and not in Cyp2e1-null mice. Metabolomic analysis demonstrated that production of isoniazid metabolites was elevated in wild-type mice along with a higher abundance of bile acids, bile acid metabolites, carnitine and carnitine derivatives; these were not observed in Cyp2e1-null mice. In addition, the enzymes responsible for bile acid synthesis were decreased and proteins involved in bile acid transport were significantly increased in wild-type mice. Lastly, treatment of targeted isoniazid metabolites to wild-type mice led to similar changes in cholesterol, triglycerides and free fatty acids. These findings suggest that while CYP2E1 is not involved in isoniazid-induced hepatotoxicity, while an isoniazid metabolite might play a role in isoniazid-induced cholestasis through enhancement of bile acid accumulation and mitochondria β-oxidation. Published by Elsevier Inc.

  6. Neutrophil Resolvin E1 Receptor Expression and Function in Type 2 Diabetes.

    Science.gov (United States)

    Freire, Marcelo O; Dalli, Jesmond; Serhan, Charles N; Van Dyke, Thomas E

    2017-01-15

    Unresolved inflammation is key in linking metabolic dysregulation and the immune system in type 2 diabetes. Successful regulation of acute inflammation requires biosynthesis of specialized proresolving lipid mediators, such as E-series resolvin (RvE) 1, and activation of cognate G protein-coupled receptors. RvE1 binds to leukotriene B4 (BLT-1) on neutrophils and to ERV-1/ChemR23 on monocyte/macrophages. We show novel actions of RvE1 and expression patterns of neutrophil receptors in type 2 diabetes. Neutrophils from healthy subjects express functional BLT-1, low levels of minimally functional ERV-1, and inversed coexpression when compared to neutrophils from type 2 diabetes subjects. Stimulation with TNF-α or LPS increased the expression of ERV-1 by healthy and diabetic neutrophils. RvE1 counteracted LPS and TNF-α induction of ERV-1 overexpression and endogenous diabetic overexpression, activating phagocytosis and resolution signals. Functional ERV-1 was determined by phosphorylation of the signaling protein ribosomal S6. Receptor-antagonism experiments revealed that the increase in phosphorylation of ribosomal S6 was mediated by BLT-1 in healthy subject neutrophils and by ERV-1 in diabetes. Metabololipidomics reveal a proinflammatory profile in diabetic serum. Cell phagocytosis is impaired in type 2 diabetes and requires RvE1 for activation. The dose of RvE1 required to activate resolution signals in type 2 diabetic neutrophils was significantly higher than in healthy controls. RvE1 rescues the dysregulation seen on neutrophil receptor profile and, following a therapeutic dosage, activates phagocytosis and resolution signals in type 2 diabetes. These findings reveal the importance of resolution receptors in health, disease, and dysregulation of inflammation in type 2 diabetes.

  7. DDX3 regulates cell growth through translational control of cyclin E1.

    Science.gov (United States)

    Lai, Ming-Chih; Chang, Wen-Cheng; Shieh, Sheau-Yann; Tarn, Woan-Yuh

    2010-11-01

    DDX3 belongs to the DEAD box family of RNA helicases, but the details of its biological function remain largely unclear. Here we show that knockdown of DDX3 expression impedes G(1)/S-phase transition of the cell cycle. To know how DDX3 may act in cell cycle control, we screened for cellular mRNA targets of DDX3. Many of the identified DDX3 targets encoded cell cycle regulators, including G(1)/S-specific cyclin E1. DDX3 depletion specifically downregulates translation of cyclin E1 mRNA. Moreover, our data suggest that DDX3 participates in translation initiation of targeted mRNAs as well as in cell growth control via its RNA helicase activity. Consistent with these findings, we show that in the temperature-sensitive DDX3 mutant hamster cell line tsET24, cyclin E1 expression is downregulated at a nonpermissive temperature that inactivates mutant DDX3. Taken together, our results indicate that DDX3 is critical for translation of cyclin E1 mRNA, which provides an alternative mechanism for regulating cyclin E1 expression during the cell cycle.

  8. Cooperative effects for CYP2E1 differ between styrene and its metabolites

    Science.gov (United States)

    Hartman, Jessica H.; Boysen, Gunnar; Miller, Grover P.

    2014-01-01

    Cooperative interactions are frequently observed in the metabolism of drugs and pollutants by cytochrome P450s; nevertheless, the molecular determinants for cooperativity remain elusive. Previously, we demonstrated that steady-state styrene metabolism by CYP2E1 exhibits positive cooperativity.We hypothesized that styrene metabolites have lower affinity than styrene toward CYP2E1 and limited ability to induce cooperative effects during metabolism. To test the hypothesis, we determined the potency and mechanism of inhibition for styrene and its metabolites toward oxidation of 4-nitrophenol using CYP2E1 Supersomes® and human liver microsomes.Styrene inhibited the reaction through a mixed cooperative mechanism with high affinity for the catalytic site (67 μM) and lower affinity for the cooperative site (1100 μM), while increasing substrate turnover at high concentrations. Styrene oxide and 4-vinylphenol possessed similar affinity for CYP2E1. Styrene oxide behaved cooperatively like styrene, but 4-vinylphenol decreased turnover at high concentrations. Styrene glycol was a very poor competitive inhibitor. Among all compounds, there was a positive correlation with binding and hydrophobicity.Taken together, these findings for CYP2E1 further validate contributions of cooperative mechanisms to metabolic processes, demonstrate the role of molecular structure on those mechanisms and underscore the potential for heterotropic cooperative effects between different compounds. PMID:23327532

  9. Silencing E1A mRNA by RNA interference inhibits adenovirus replication.

    Science.gov (United States)

    Chung, Y-S; Kim, M-K; Lee, W-J; Kang, C

    2007-01-01

    The adenovirus family contains 51 human serotypes, and most human adenoviruses cause widespread respiratory tract infections. Adenovirus infections can result in severe complications in some cases, such as in adenovirus type 11 infection in immunocompromised patients. However, effective treatment methods for adenovirus infections are currently unavailable. This prompted the search for antiviral agents effective against adenovirus infections. In the present study, adenovirus E1A was targeted by RNA interference (RNAi) using synthetic small interfering RNAs (siRNAs) in an attempt to inhibit viral replication, since adenovirus E1A proteins are known to be involved in the transcriptional activation of the viral and cellular genes necessary for controlling the cell cycle and viral replication. The results indicated that the siRNAs effectively reduced the amount of adenovirus E1A mRNA and the levels of replicative intermediates. Additionally, siRNA-mediated gene silencing inhibited adenovirus replication by suppressing the E1A mRNA. These results suggest that the RNAi-mediated targeting of adenovirus E1A may have a potentially therapeutic effect in controlling adenovirus infections.

  10. 2-Carboxyquinoxalines kill mycobacterium tuberculosis through noncovalent inhibition of DprE1.

    Science.gov (United States)

    Neres, João; Hartkoorn, Ruben C; Chiarelli, Laurent R; Gadupudi, Ramakrishna; Pasca, Maria Rosalia; Mori, Giorgia; Venturelli, Alberto; Savina, Svetlana; Makarov, Vadim; Kolly, Gaelle S; Molteni, Elisabetta; Binda, Claudia; Dhar, Neeraj; Ferrari, Stefania; Brodin, Priscille; Delorme, Vincent; Landry, Valérie; de Jesus Lopes Ribeiro, Ana Luisa; Farina, Davide; Saxena, Puneet; Pojer, Florence; Carta, Antonio; Luciani, Rosaria; Porta, Alessio; Zanoni, Giuseppe; De Rossi, Edda; Costi, Maria Paola; Riccardi, Giovanna; Cole, Stewart T

    2015-03-20

    Phenotypic screening of a quinoxaline library against replicating Mycobacterium tuberculosis led to the identification of lead compound Ty38c (3-((4-methoxybenzyl)amino)-6-(trifluoromethyl)quinoxaline-2-carboxylic acid). With an MIC99 and MBC of 3.1 μM, Ty38c is bactericidal and active against intracellular bacteria. To investigate its mechanism of action, we isolated mutants resistant to Ty38c and sequenced their genomes. Mutations were found in rv3405c, coding for the transcriptional repressor of the divergently expressed rv3406 gene. Biochemical studies clearly showed that Rv3406 decarboxylates Ty38c into its inactive keto metabolite. The actual target was then identified by isolating Ty38c-resistant mutants of an M. tuberculosis strain lacking rv3406. Here, mutations were found in dprE1, encoding the decaprenylphosphoryl-d-ribose oxidase DprE1, essential for biogenesis of the mycobacterial cell wall. Genetics, biochemical validation, and X-ray crystallography revealed Ty38c to be a noncovalent, noncompetitive DprE1 inhibitor. Structure-activity relationship studies generated a family of DprE1 inhibitors with a range of IC50's and bactericidal activity. Co-crystal structures of DprE1 in complex with eight different quinoxaline analogs provided a high-resolution interaction map of the active site of this extremely vulnerable target in M. tuberculosis.

  11. Activation of the human beta interferon gene by the adenovirus type 12 E1B gene

    Energy Technology Data Exchange (ETDEWEB)

    Shiroki, K.; Toth, M.

    1988-01-01

    The transcription of endogenous beta interferon mRNA was activated in human embryo kidney (HEK) cells infected with adenovirus 12 (Ad12) but was activated only inefficiently or not at all in HEK cells infected with Ad5 and rc-1 (Ad5 dl312 containing the Ad12 E1A region). The analysis with Ad12 mutants showed that Ad12 E1B products, especially the 19K protein, were important for the expression of the endogenous beta interferon gene and Ad12 E1A products were not involved in the expression. The expression of exogeneously transfected pIFN-CAT (a hybrid plasmid having the human beta interferon promoter fused with the CAT gene) was activated in HEK and chicken embryo fibroblast (CEF) cells infected with either Ad12 or Ad5. The analysis of cotransfection of CEF cells with pIFN-CAT and plasmids containing fragments of Ad12 or Ad5 DNA showed that Ad12 or Ad5 E1B (possibly the 19K protein) was and E1A was not involved in the expression of the exogenous pIFN-CAT.

  12. E1B and E4 oncoproteins of adenovirus antagonize the effect of apoptosis inducing factor

    Energy Technology Data Exchange (ETDEWEB)

    Turner, Roberta L. [Department of Microbiology and Immunology, Wake Forest School of Medicine, Winston-Salem, NC 27157 (United States); Wilkinson, John C., E-mail: john.wilkinson@ndsu.edu [Department of Biochemistry, Wake Forest School of Medicine, Winston-Salem, NC 27157 (United States); Ornelles, David A., E-mail: ornelles@wakehealth.edu [Department of Microbiology and Immunology, Wake Forest School of Medicine, Winston-Salem, NC 27157 (United States)

    2014-05-15

    Adenovirus inundates the productively infected cell with linear, double-stranded DNA and an abundance of single-stranded DNA. The cellular response to this stimulus is antagonized by the adenoviral E1B and E4 early genes. A mutant group C adenovirus that fails to express the E1B-55K and E4ORF3 genes is unable to suppress the DNA-damage response. Cells infected with this double-mutant virus display significant morphological heterogeneity at late times of infection and frequently contain fragmented nuclei. Nuclear fragmentation was due to the translocation of apoptosis inducing factor (AIF) from the mitochondria into the nucleus. The release of AIF was dependent on active poly(ADP-ribose) polymerase-1 (PARP-1), which appeared to be activated by viral DNA replication. Nuclear fragmentation did not occur in AIF-deficient cells or in cells treated with a PARP-1 inhibitor. The E1B-55K or E4ORF3 proteins independently prevented nuclear fragmentation subsequent to PARP-1 activation, possibly by altering the intracellular distribution of PAR-modified proteins. - Highlights: • E1B-55K or E4orf3 prevents nuclear fragmentation. • Nuclear fragmentation requires AIF and PARP-1 activity. • Adenovirus DNA replication activates PARP-1. • E1B-55K or E4orf3 proteins alter the distribution of PAR.

  13. Purification and application of C-terminally truncated hepatitis C virus E1 proteins expressed in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    Jing Liu; Li-Xin Zhu; Yu-Ying Kong; Guang-Di Li; Yuan Wang

    2005-01-01

    AIM: To explore the possibility of expressing hepatitis C virus (HCV) envelope protein 1 (E1) in Escherichia coli(E.coli)and to test the purified recombinant E1 proteins for clinical and research applications. METHODS: C-terminally trunczated E1 fragments were expressed in E. coli as hexa-histidine-tagged fusion proteins. The expression products were purified under denaturing conditions using immobilized-metal affinity chromatography. Purified E1 proteins were used to immunize rabbits. Rabbit anti-sera thus obtained were reacted with both E. coli- and mammalian cell-expressed E1 glycoproteins as detected by Western blot.RESULTS: Full-length E1 protein proved difficult to express in E. coli. C-terminally truncated E1 was successfully expressed in E. coli as hexa-histidine-tagged recombinant fusion protein and was purified under denaturing conditions on Ni2+-NTA agarose. Rabbit anti-sera raised against purified recombinant E1 specifically reacted with mammalian cell-expressed E1 giycoproteins in Western blot. Furthermore, E. coli-derived E1 protein was able to detect animal antibodies elicited by E1-based DNA immunization.CONCLUSION: These results demonstrate that the prokaryotically expressed E1 proteins share identical epitopes with eukaryotically expressed E1 glycoprotein. The E. coli-derived E1 proteins and corresponding antisera can become useful tools in anti-HCV vaccine research.

  14. Calcification of MC3T3-E1 cells on titanium and zirconium.

    Science.gov (United States)

    Umezawa, Takayuki; Chen, Peng; Tsutsumi, Yusuke; Doi, Hisashi; Ashida, Maki; Suzuki, Shoichi; Moriyama, Keiji; Hanawa, Takao

    2015-01-01

    To confirm similarity of hard tissue compatibility between titanium and zirconium, calcification of MC3T3-E1 cells on titanium and zirconium was evaluated in this study. Mirror-polished titanium (Ti) and zirconium (Zr) disks and zirconium-sputter deposited titanium (Zr/Ti) were employed in this study. The surface of specimens were characterized using scanning electron microscopy and X-ray diffraction. Then, the cellular proliferation, differentiation and calcification of MC3T3-E1 cells on specimens were investigated. The surface of Zr/Ti was much smoother and cleaner than those of Ti and Zr. The proliferation of the cell was the same among three specimens, while the differentiation and calcification on Zr/Ti were faster than those on Ti and Zr. Therefore, Ti and Zr showed the identical hard tissue compatibility according to the evaluation with MC3T3-E1 cells. Sputter deposition may improve cytocompatibility.

  15. Double ion beams and accelerated ions observed by SWIDs on Chang'E-1

    Science.gov (United States)

    Kong, Linggao; Zhang, Aibing; Wang, Shijin

    2012-07-01

    Chang'E-1 is the first Chinese lunar exploration spacecraft launced in 2007. Solar Wind Ion Detectors(SWIDs) are two scientific instruments on Chang'E-1 to measure the solar wind and the plasma environment around the Moon. Some cases of double ion beams in solar wind were observed by SWIDs. These double ion beams present double proton beams with a single alpha beam. Another insteresting phenomenon observed by SWIDs is the accelerated ions when the spacecraft was over the magnetic anomalies. The magnetic anomaly may induce mini-magnetosphere, which was confirmed both by simulation and measurement of some other lunar exploration spacecrafts. The accelerated ions observed by Chang'E-1 may be the result of the interaction of the solar wind and the mini-magnetosphere of the Moon.

  16. Influence of Lactobacillus E1 on the Storage Stability in Emulsion Immobilization

    Institute of Scientific and Technical Information of China (English)

    SUN Xin; CHEN Xiguang; LIU Chengsheng; CHA Dongsu; PENG Hongni

    2009-01-01

    A coacervation method with double emulsion strategy (w/o/w) was used to prepare immobilized Lactobacillus E1. Diatomite was chosen as the carrier for bacteria. Sodium alginate,dextrin and gelatin were used as protective solutes for the preservation of Lactobacillus E1 and their effects on the storage viability during storage were discussed. The influence of storage temperature on the storage viability was also examined. The results show that high bacteria viable count over 109 cfu/g for an extended shelf life of 37 d can be achieved with 2% sodium alginate, 5% dextrin and 4% gelatin as protective solutes, at 10 ℃ of the storage temperature. This immobilized Lactobacillus E1 has potential use as functional food ingredient for both human dairy food and animal feedstuff.

  17. Melanoma cultures show different susceptibility towards E1A-, E1B-19 kDa- and fiber-modified replication-competent adenoviruses.

    Science.gov (United States)

    Schmitz, M; Graf, C; Gut, T; Sirena, D; Peter, I; Dummer, R; Greber, U F; Hemmi, S

    2006-06-01

    Replicating adenovirus (Ad) vectors with tumour tissue specificity hold great promise for treatment of cancer. We have recently constructed a conditionally replicating Ad5 AdDeltaEP-TETP inducing tumour regression in a xenograft mouse model. For further improvement of this vector, we introduced four genetic modifications and analysed the viral cytotoxicity in a large panel of melanoma cell lines and patient-derived melanoma cells. (1) The antiapoptotic gene E1B-19 kDa (Delta19 mutant) was deleted increasing the cytolytic activity in 18 of 21 melanoma cells. (2) Introduction of the E1A 122-129 deletion (Delta24 mutant), suggested to attenuate viral replication in cell cycle-arrested cells, did not abrogate this activity and increased the cytolytic activity in two of 21 melanoma cells. (3) We inserted an RGD sequence into the fiber to extend viral tropism to alphav integrin-expressing cells, and (4) swapped the fiber with the Ad35 fiber (F35) enhancing the tropism to malignant melanoma cells expressing CD46. The RGD-fiber modification strongly increased cytolysis in all of the 11 CAR-low melanoma cells. The F35 fiber-chimeric vector boosted the cytotoxicity in nine of 11 cells. Our results show that rational engineering additively enhances the cytolytic potential of Ad vectors, a prerequisite for the development of patient-customized viral therapies.

  18. Inhibitory Potency of 4-Carbon Alkanes and Alkenes toward CYP2E1 Activity

    Science.gov (United States)

    Hartman, Jessica H.; Miller, Grover P.; Boysen, Gunnar

    2016-01-01

    CYP2E1 has been implicated in the bioactivation of many small molecules into reactive metabolites which form adducts with proteins and DNA, and thus a better understanding of the molecular determinants of its selectivity are critical for accurate toxicological predictions. In this study, we determined the potency of inhibition of human CYP2E1 for various 4-carbon alkanes, alkenes and alcohols. In addition, known CYP2E1 substrates and inhibitors including 4-methylpyrazole, aniline, and dimethylnitrosamine were included to determine their relative potencies. Of the 1,3-butadiene-derived metabolites studied, 3,4-epoxy-1-butene was the strongest inhibitor with an IC50 of 110 μM compared to 1700 μM and 6600 μM for 1,2-butenediol and 1,2:3,4-diepoxybutane, respectively. Compared to known inhibitors, inhibitory potency of 3,4-epoxy-1-butene is between 4-methylpyrazole (IC50 = 1.8 μM) and dimethylnitrosamine (IC50 = 230 μM). All three butadiene metabolites inhibit CYP2E1 activity through a simple competitive mechanism. Among the 4-carbon compounds studied, the presence and location of polar groups seems to influence inhibitory potency. To further examine this notion, the investigation was extended to include structurally and chemically similar analogs, including propylene oxide and various butane alcohols. Those results demonstrated preferential recognition of CYP2E1 toward the type and location of polar and hydrophobic structural elements. Taken together, CYP2E1 metabolism may be modified in vivo by exposure to 4-carbon compounds, such as drugs, and nutritional constituents, a finding that highlights the complexity of exposure to mixtures. PMID:24561005

  19. Propolis protects CYP 2E1 enzymatic activity and oxidative stress induced by carbon tetrachloride.

    Science.gov (United States)

    Bhadauria, Monika; Nirala, Satendra Kumar; Shukla, Sangeeta

    2007-08-01

    Induction of CYP 2E1 by carbon tetrachloride (CCl(4)) is one of the central pathways by which CCl(4) generates oxidative stress in hepatocytes. Experimental liver injury was induced in rats by CCl(4) to determine toxicological actions on CYP 2E1 by microsomal drug metabolizing enzymes. In this report, ethanolic extract of propolis at a dose of 200 mg/kg (po) was used after 24 h of toxicant administration to validate its protective potential. Intraperitoneal injection of CCl(4) (1.5 ml/kg) induced hepatotoxicity after 24 h of its administration that was associated with elevated malonyldialdehyde (index of lipid peroxidation), lactate dehydrogenase and gamma-glutamyl transpeptidase release (index of a cytotoxic effect). Hepatic microsomal drug metabolizing enzymes of CYP 2E1 showed sharp depletion as assessed by estimating aniline hydroxylase and amidopyrine N-demethylase activity after CCl(4) exposure. Toxic effect of CCl(4) was evident on CYP 2E1 activity by increased hexobarbitone induced sleep time and bromosulphalein retention. Propolis extract showed significant improvement in the activity of both enzymes and suppressed toxicant induced increase in sleep time and bromosulphalein retention. Choleretic activity of liver did not show any sign of toxicity after propolis treatment at a dose of 200 mg/kg (id). Histopathological evaluation of the liver revealed that propolis reduced the incidence of liver lesions including hepatocyte swelling and lymphocytic infiltrations induced by CCl(4). Electron microscopic observations also showed improvement in ultrastructure of liver and substantiated recovery in biochemical parameters. Protective activity of propolis at 200 mg/kg dose was statistically compared with positive control silymarin (50 mg/kg, po), a known hepatoprotective drug seems to be better in preventing hepatic CYP 2E1 activity deviated by CCl(4). These results lead us to speculate that propolis may play hepatoprotective role via improved CYP 2E1 activity and

  20. p53/E1b58kDa complex regulates adenovirus replication.

    Science.gov (United States)

    Ridgway, P J; Hall, A R; Myers, C J; Braithwaite, A W

    1997-10-27

    We have explored a role for the adenovirus (Ad5) E1b58kDa/p53 protein complex in adenovirus replication. This was done by using virus mutants containing different defects in the E1b58kDa gene and cell lines that express either a wild-type p53 protein or a mutant p53 protein. We find that infection of wild-type p53-containing cells with wild-type Ad5 causes a shutoff of p53 and alpha-actin protein synthesis by distinct mechanisms, but neither occurs in mutant p53 cells. Our data also indicate that the shutoff is dependent on formation of the p53/E1b complex and may also involve another virus protein, E4ORF6. Following from these observations we asked whether failure to form the complex resulted in impaired adenovirus replication. Our experiments showed that neither wild-type Ad5 nor the E1b mutant dl338 could replicate in cells expressing a mutant p53 protein, but that wild-type adenovirus replicated well in wild-type p53-expressing cells. Collectively, our data suggest that the interaction between p53 and the E1b58kDa protein is necessary for efficient adenovirus replication. This is the first time such a direct link between the complex and virus replication has been demonstrated. These data raise serious questions about the usefulness of E1b-defective viruses in tumor therapy.

  1. Functional characterisation of an engineered multidomain human P450 2E1 by molecular Lego.

    Science.gov (United States)

    Fairhead, Michael; Giannini, Silva; Gillam, Elizabeth M J; Gilardi, Gianfranco

    2005-12-01

    The human cytochrome P450s constitute an important family of monooxygenase enzymes that carry out essential roles in the metabolism of endogenous compounds and foreign chemicals. We present here results of a fusion between a human P450 enzyme and a bacterial reductase that for the first time is shown does not require the addition of lipids or detergents to achieve wild-type-like activities. The fusion enzyme, P450 2E1-BMR, contains the N-terminally modified residues 22-493 of the human P450 2E1 fused at the C-terminus to residues 473-1049 of the P450 BM3 reductase (BMR). The P450 2E1-BMR enzyme is active, self-sufficient and presents the typical marker activities of the native human P450 2E1: the hydroxylation of p-nitrophenol (KM=1.84+/-0.09 mM and kcat of 2.98+/-0.04 nmol of p-nitrocatechol formed per minute per nanomole of P450) and chlorzoxazone (KM=0.65+/-0.08 mM and kcat of 0.95+/-0.10 nmol of 6-hydroxychlorzoxazone formed per minute per nanomole of P450). A 3D model of human P450 2E1 was generated to rationalise the functional data and to allow an analysis of the surface potentials. The distribution of charges on the model of P450 2E1 compared with that of the FMN domain of BMR provides the ground for the understanding of the interaction between the fused domains. The results point the way to successfully engineer a variety of catalytically self-sufficient human P450 enzymes for drug metabolism studies in solution.

  2. DDX3 Regulates Cell Growth through Translational Control of Cyclin E1▿ †

    OpenAIRE

    2010-01-01

    DDX3 belongs to the DEAD box family of RNA helicases, but the details of its biological function remain largely unclear. Here we show that knockdown of DDX3 expression impedes G1/S-phase transition of the cell cycle. To know how DDX3 may act in cell cycle control, we screened for cellular mRNA targets of DDX3. Many of the identified DDX3 targets encoded cell cycle regulators, including G1/S-specific cyclin E1. DDX3 depletion specifically downregulates translation of cyclin E1 mRNA. Moreover, ...

  3. Thionin-D4E1 chimeric protein protects plants against bacterial infections

    Energy Technology Data Exchange (ETDEWEB)

    Stover, Eddie W; Gupta, Goutam; Hao, Guixia

    2017-08-08

    The generation of a chimeric protein containing a first domain encoding either a pro-thionon or thionin, a second domain encoding D4E1 or pro-D4E1, and a third domain encoding a peptide linker located between the first domain and second domain is described. Either the first domain or the second domain is located at the amino terminal of the chimeric protein and the other domain (second domain or first domain, respectively) is located at the carboxyl terminal. The chimeric protein has antibacterial activity. Genetically altered plants and their progeny expressing a polynucleotide encoding the chimeric protein resist diseases caused by bacteria.

  4. Improvements and Testing Practical Expressions for Photon Strength Functions of E1 Gamma-Transitions

    CERN Document Server

    Plujko, Vladimir; Kadenko, Igor; Solodovnyk, Kateryna

    2016-01-01

    Analytical expression for the E1 photon strength functions (PSF) is modified to account for the low-energy enhancement due to nuclear structure effects (presence of low-energy state (LES)). A closed-form expression of the E1 PSF function includes response of two nuclear states: LES and giant diplole resonance (GDR). Expression for the nuclear response function on electromagnetic field is based on a model of excitation of two coupled damped states. These approach is tested for different data sets for spherical nuclei. Impact on PSF shape of coupling between LES and GDR excitations is considered.

  5. E1A genes of adenovirus type 2 and type 5 are expressed at different levels

    DEFF Research Database (Denmark)

    Moritz, Constanze; Dobbelstein, Matthias

    2006-01-01

    Adenoviruses are an extensively studied system for modeling oncogenesis and for experimental cancer therapy. The most commonly analyzed virus types are 2 and 5, and little distinction has been made between them in past studies. Adenoviruses used for therapeutic purposes are frequently hybrids...... region. We found that the hybrid viruses replicated with considerably lower efficiency than their type 5 counterparts in H1299 cells (dl309:WtD = 3-4, dl338:dl1520 > 10). Moreover, adenovirus type 2 E1A expression from the hybrid viruses was strongly reduced in comparison to adenovirus type 5 E1A...

  6. CYP2E1-dependent elevation of serum cholesterol, triglycerides, and hepatic bile acids by isoniazid

    Energy Technology Data Exchange (ETDEWEB)

    Cheng, Jie; Krausz, Kristopher W. [Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 (United States); Li, Feng; Ma, Xiaochao [Department of Pharmacology, Toxicology and Therapeutics, The University of Kansas Medical Center, 4089 KLSIC, MS 1018, 3901 Rainbow Boulevard, Kansas City, KS 66160 (United States); Gonzalez, Frank J., E-mail: fjgonz@helix.nih.gov [Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 (United States)

    2013-01-15

    Isoniazid is the first-line medication in the prevention and treatment of tuberculosis. Isoniazid is known to have a biphasic effect on the inhibition–induction of CYP2E1 and is also considered to be involved in isoniazid-induced hepatotoxicity. However, the full extent and mechanism of involvement of CYP2E1 in isoniazid-induced hepatotoxicity remain to be thoroughly investigated. In the current study, isoniazid was administered to wild-type and Cyp2e1-null mice to investigate the potential toxicity of isoniazid in vivo. The results revealed that isoniazid caused no hepatotoxicity in wild-type and Cyp2e1-null mice, but produced elevated serum cholesterol and triglycerides, and hepatic bile acids in wild-type mice, as well as decreased abundance of free fatty acids in wild-type mice and not in Cyp2e1-null mice. Metabolomic analysis demonstrated that production of isoniazid metabolites was elevated in wild-type mice along with a higher abundance of bile acids, bile acid metabolites, carnitine and carnitine derivatives; these were not observed in Cyp2e1-null mice. In addition, the enzymes responsible for bile acid synthesis were decreased and proteins involved in bile acid transport were significantly increased in wild-type mice. Lastly, treatment of targeted isoniazid metabolites to wild-type mice led to similar changes in cholesterol, triglycerides and free fatty acids. These findings suggest that while CYP2E1 is not involved in isoniazid-induced hepatotoxicity, while an isoniazid metabolite might play a role in isoniazid-induced cholestasis through enhancement of bile acid accumulation and mitochondria β-oxidation. -- Highlights: ► Isoniazid metabolites were elevated only in wild-type mice. ► Isoniazid caused no hepatotoxicity in wild-type and Cyp2e1-null mice. ► Isoniazid elevated serum cholesterol and triglycerides, and hepatic bile acids. ► Bile acid transporters were significantly decreased in isoniazid-treated mice.

  7. A regulatory variant in CYP2E1 affects the risk of lung squamous cell carcinoma.

    Science.gov (United States)

    Cao, Lei; Lin, Jia; He, Bing; Wang, Hongge; Rao, Juan; Liu, Yingwen; Zhang, Xuemei

    2014-01-01

    Cytochrome P450 2E1 (CYP2E1), an ethanol-inducible enzyme, has been shown to metabolically activate various carcinogens, which is critical for the development of cancers. It has been demonstrated that CYP2E1 polymorphisms alter the transcriptional activity. However, studies on the association between CYP2E1 -1239G>C polymorphism and non-small cell lung cancer have reported conflicting results. Thus, the gain of the present study was to investigate whether CYP2E1 -1239G>C polymorphism is associated with the development of non-small cell lung cancer in Chinese population. A case-control study was conducted in which CYP2E1 -1239G>C polymorphism was analyzed in 526 Chinese patients with non-small cell lung cancer and 526 age-matched healthy controls by polymerase chain reaction-restriction fragment length polymorphism. Odds ratios were estimated by multivariate logistic regression. A meta-analysis was conducted to evaluate the association of CYP2E1 -1239G>C polymorphism with the risk of lung cancer in Chinese population by calculating pooled odds ratio (OR). For CYP2E1 -1239G>C polymorphism, -1239C allele carriers (OR = 0.67; 95% confidence interval (CI) = 0.51-0.87; P = 0.002) were associated with a decreased risk of non-small cell lung cancer when compared with -1239GG homozygotes. In the group analyses by pathological types, for lung squamous cell carcinoma and other types, the ORs of the C allele carriers were 0.60 (95% CI = 0.41-0.88; P = 0.009) and 0.54 (95% CI = 0.30-0.99; P = 0.045). In the group analysis of smoking status, the OR for the -1239C allele-containing genotype was higher than that for -1239GG genotype (OR = 0.57; 95% CI = 0.40-0.81; P = 0.002) among smokers, but not among nonsmokers. Moreover, when the risk associated with CYP2E1 polymorphism was further evaluated within strata of C polymorphism and the risk of non-small cell lung cancer. Meta-analysis data also showed that the carriers of CYP2E1 -1239C allele

  8. Geologic investigation and mapping of the Sinus Iridum quadrangle from Clementine, SELENE, and Chang’e-1 data

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The objectives of lunar satellite remote sensing are to study lunar surface characteristics, inner structure, and its evolution history. The contents of TiO 2 and FeO are assessed from Clementine UV/VIS data for Sinus Iridum. The geologic stratigraphic units and crates are interpreted visually based on SELENE Terrain Camera (TC) images and the spatial resolution of which is up to 10 m. And the geologic ages of different stratigraphic units are calculated by the crater size-frequency distributions measurements. The gravity anomaly is generated from SELENE gravity model (SGM90d) to show its difference from Mare Imbrium. Furthermore, the thickness of lunar regolith is also derived from microwave radiometer data of Chang’e-1 satellite. Integrating these results, it shows that the Sinus Iridum is different from the Mare Imbrium in inner structure and surface sedimentation. And its history of subsidence, deposition, volcanism, and impact is described. It makes sense to the future soft-landing and sampling at potential Sinus Iridum by remote sensing geologic analysis.

  9. PDH-E1alpha dephosphorylation and activation in human skeletal muscle during exercise

    DEFF Research Database (Denmark)

    Pilegaard, Henriette; Birk, Jesper Bratz; Sacchetti, Massimo;

    2006-01-01

    extensor exercise at moderate intensity. During the 4-h resting period, activity of PDH in the active form (PDHa) did not change in either trial, yet phosphorylation of PDH-E1a site 1 (PDH-P1) and site 2 (PDH-P2) was elevated in the intralipid compared with the control trial. PDHa activity increased during...

  10. US NDC Modernization Iteration E1 Prototyping Report: User Interface Framework

    Energy Technology Data Exchange (ETDEWEB)

    Lober, Randall R. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2014-12-01

    During the first iteration of the US NDC Modernization Elaboration phase (E1), the SNL US NDC modernization project team completed an initial survey of applicable COTS solutions, and established exploratory prototyping related to the User Interface Framework (UIF) in support of system architecture definition. This report summarizes these activities and discusses planned follow-on work.

  11. Methylglyoxal induces oxidative stress and mitochondrial dysfunction in osteoblastic MC3T3-E1 cells.

    Science.gov (United States)

    Suh, K S; Choi, E M; Rhee, S Y; Kim, Y S

    2014-02-01

    Methylglyoxal is a reactive dicarbonyl compound produced by glycolytic processing and identified as a precursor of advanced glycation end products. The elevated methylglyoxal levels in patients with diabetes are believed to contribute to diabetic complications, including bone defects. The objective of this study was to evaluate the effect of methylglyoxal on the function of osteoblastic MC3T3-E1 cells. The data indicated that methylglyoxal decreased osteoblast differentiation and induced osteoblast cytotoxicity. Pretreatment of MC3T3-E1 cells with aminoguanidine (a carbonyl scavenger), Trolox (an antioxidant), and cyclosporin A (a blocker of the mitochondrial permeability transition pore) prevented methylglyoxal-induced cytotoxicity in MC3T3-E1 cells. However, BAPTA/AM (an intracellular Ca(2+) chelator) and dantrolene (an inhibitor of endoplasmic reticulum Ca(2+) release) did not reverse the cytotoxic effect of methylglyoxal. Methylglyoxal increased the formation of intracellular reactive oxygen species, mitochondrial superoxide, and cardiolipin peroxidation in osteoblastic MC3T3-E1 cells. Methylglyoxal also decreased the mitochondrial membrane potential and intracellular ATP and nitric oxide levels, suggesting that carbonyl stress-induced loss of mitochondrial integrity contributes to the cytotoxicity of methylglyoxal. Furthermore, the results demonstrated that methylglyoxal induced protein adduct formation, inactivation of glyoxalase I, and activation of glyoxalase II. Aminoguanidine reversed all aforementioned effects of methylglyoxal. Taken together, these data support the notion that high methylglyoxal concentrations have detrimental effects on osteoblasts through a mechanism involving oxidative stress and mitochondrial dysfunction.

  12. 26 CFR 301.6511(e)-1 - Special rules applicable to manufactured sugar.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 18 2010-04-01 2010-04-01 false Special rules applicable to manufactured sugar... Assessment and Collection § 301.6511(e)-1 Special rules applicable to manufactured sugar. (a) Use as... the person entitled thereto. Such right accrues as of the date the manufactured sugar, or article...

  13. US NDC Modernization Iteration E1 Prototyping Report: Processing Control Framework

    Energy Technology Data Exchange (ETDEWEB)

    Prescott, Ryan [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Hamlet, Benjamin R. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2014-12-01

    During the first iteration of the US NDC Modernization Elaboration phase (E1), the SNL US NDC modernization project team developed an initial survey of applicable COTS solutions, and established exploratory prototyping related to the processing control framework in support of system architecture definition. This report summarizes these activities and discusses planned follow-on work.

  14. US NDC Modernization Iteration E1 Prototyping Report: Common Object Interface

    Energy Technology Data Exchange (ETDEWEB)

    Lewis, Jennifer E. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Hess, Michael M. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2014-12-01

    During the first iteration of the US NDC Modernization Elaboration phase (E1), the SNL US NDC modernization project team completed an initial survey of applicable COTS solutions, and established exploratory prototyping related to the Common Object Interface (COI) in support of system architecture definition. This report summarizes these activities and discusses planned follow-on work.

  15. Transcriptional activation by the E1A regions of adenovirus types 40 and 41

    NARCIS (Netherlands)

    Loon, A.E. van; Gilardi, P.; Perricaudet, M.; Rozijn, Th. H.; Sussenbach, J.S.

    1987-01-01

    In order to establish whether the poor growth of the two fastidious adenoviruses types 40 and 41 (Ad40 and Ad41) in HeLa cells is due to a reduced trans-activation by the early region to (E1A), we have determined the trans-activating effect of this region on the expression of the chloramphenicol ace

  16. Two-step method for curing Escherichia coli of ColE1-derived plasmids

    DEFF Research Database (Denmark)

    Hove-Jensen, Bjarne

    2008-01-01

    To cure Escherichia coli for plasmids derived from the ColE1 replicon advantage is taken of the fact that maintenance of this replicon requires a wild-type allele of polA, encoding DNA polymerase I. Curing is achieved by cotransduction of a mutant polA allele with metE::Tn10, fadAB::Tn10 or other...

  17. E1A,E1B double-restricted adenovirus enhances the cytotoxicity and antitumor activity of gemcitabine to renal cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    WANG Hua; Makoto Satoh; CHEN Gui-ping; LI De-chuan; Hirofumi Hamada; Yoichi Arai

    2011-01-01

    Background Our previous studies have demonstrated potent oncolysis efficacy of the E1A,E1B double-restricted replication-competent oncolytic adenovirus AxdAdB-3 for treatment of bladder cancer. Here,we reported the feasibility and efficacy of AxdAdB-3 alone,or in combination with gemcitabine for treating renal cell carcinoma.Methods Cytopathic effects of AxdAdB-3 were evaluated in human renal cell carcinoma cell lines TOS-1,TOS-2,TOS-3,TOS-3LN,SMKT-R3,SMKT-R4 and ACHN,and in normal human renal proximal tubule epithelial cells (RPTEC).AxdAdB-3 induced down-regulation of the cell cycle was determined by flow cytometry. Combination therapies of AxdAdB-3 with gemcitabine were evaluated in vitro and in vivo on subcutaneous TOS-3LN tumors in a severe combined immunodeficiency disease (SCID) mouse model.Results AxdAdB-3 was potently cytopathic against the tested most renal cell carcinoma cell lines including TOS-2,TOS-3,TOS-3LN,SMKT-R3 and SMKT-R4,while normal human RPTEC were not destroyed. AxdAdB-3 effectively induced cell cycle S-phase entry. Combined therapy of AxdAdB-3 with gemcitabine demonstrated stronger antitumor effects in vitro and in vivo compared with either AxdAdB-3 or gemcitabine alone.Conclusion AxdAdB-3 alone,or in combination with gemcitabine may be a promising strategy against renal cell carcinoma.

  18. The development of the CHANG'`E-1 lunar explorer laser altimeter

    Science.gov (United States)

    Shu, R.; Wang, J. Y.; Hu, Y. H.; Jia, J. J.

    According to Chinese fable CHANG prime E is the name of a peri who lives in the moon with a white rabbit The CHANG prime E-1 lunar explorer will be launched in April 2007 The Lunar Explorer Laser Altimeter LELA is one of the 6 payloads in Chang prime E-1 which is developed by Shanghai Institute of Technical Physics Chinese Academy of Sciences The data of LELA will be used with the Optical Imager for acquiring the three dimension image of the moon surface The LELA transmits laser pulses determines their round trip times to the surface of the moon using a time interval counter and measures ranges between CHANG prime E-1 lunar explorer and the lunar surface in the nadir direction with 5m accuracy every 1 second for 1 year s mission period The acquired range data are transformed to the topography of the moon with the aid of position and attitude data of the CHANG prime E-1 lunar explorer obtained from the ground-based tracking and on board star sensor respectively The mean distances between Chang prime E-1 and the surface of moon is 200Km The LELA utilizes a laser diode LD pumped Q-switched Nd YAG laser that has a wavelength of 1064nm a pulse width of 10ns The output beam divergence is improved to 0 6mrad by Galileo refractor-type collimator which resulted in a moon surface spot size foot print of 120m The return pulses are captured by Cassegrain-type reflector and detected by Si-APD detector The prototype began from 2003 Until now the engineering model had been finished

  19. Sonochemical synthesis of novel pyrano[3,4-e][1,3]oxazines: A green protocol.

    Science.gov (United States)

    Saleh, Tamer S; Al-Bogami, Abdullah S; Mekky, Ahmed E M; Alkhathlan, Hamad Z

    2017-05-01

    The atom-efficient and green protocol for formation of pyrano[3,4-e][1,3]oxazines utilizing dimethyl carbonate under ultrasound irradiation in a presence of KF/basic alumina was reported. We provide a novel series of pyrano[3,4-e][1,3]oxazine derivatives interesting for biological screening tests. In general, it was found that ultrasound irradiations enable the reactions to occur which could not be carried out under silent conditions. These remarkable effects appeared in sonicated reactions can be reasonably interpreted in terms of acoustic cavitation phenomenon. Structures of the products were established on analytical and spectral data. This protocol offers several advantages attain many principles of green chemistry including, save energy, atom economy, clean reactions, inexpensive green reagent and use catalysts rather than stoichiometric reagents.

  20. Introduction to the Payloads and the Initial Observation Results of Chang'E-1

    Institute of Scientific and Technical Information of China (English)

    SUN Huixian; CHEN Xiaomin; OUYANG Ziyuan; ZOU Yongliao; WU Ji; DAI Shuwu; ZHAO Baochang; SHU Rong; CHANG Jin; WANG Huanyu; ZHANG Xiaohui; REN Qiongying

    2008-01-01

    Chang'E-1, the orbiter circling the moon 200km above the moon surface, is the first Chinese Lunar exploration satellite. The satellite was successfully launched on 24th October 2007.There are 8 kinds of scientific payloads onboard, including the stereo camera, the laser altimeter, the Sagnac-based interferometer image spectrometer, the Gamma ray spectrometer, the X-ray spectrometer, the microwave radiometer, the high energy particle detector, the solar wind plasma detector and a supporting payload data management system. Chang'E-1 opened her eyes to look at the moon and took the first batch of lunar pictures after her stereo camera was switched on in 20th November 2007.Henceforth all the instruments are successfully switched on one by one. After a period of parameter adjustment and initial check out, all scientific instruments are now in their normal operating phase.In this paper, the payloads and the initial observation results are introduced.

  1. Heterologous expression of human cytochrome P450 2E1 in HepG2 cell line

    Institute of Scientific and Technical Information of China (English)

    Jian Zhuge; Ye Luo; Ying-Nian Yu

    2003-01-01

    AIM: Human cytochrome P-450 2E1 (CYP2E1) takes part in the biotransformation of ethanol, acetone, many smallmolecule substrates and volatile anesthetics. CYP2E1 is involved in chemical activation of many carcinogens,procarcinogens, and toxicants. To assess the metabolic and toxicological characteristics of CYP2E1, we cloned CYP2E1 cDNA and established a HepG2 cell line stably expressing recombinant CYP 2E1.METHODS: Human CYP2E1 cDNA was amplified with reverse transcription-polymerase chain reaction (RT-PCR)from total RNAs extracted from human liver and cloned into pGEM-T vector. The cDNA segment was identified by DNA sequencing and subcloned into a mammalian expression vector pREP9. A transgenic cell line was established by transfecting the recombinant plasmid of pREP9-CYP2E1 to HepG2 cells. The expression of CYP2E1 mRNA was validated by RT-PCR. The enzyme activity of CYP2E1 catalyzing oxidation of 4-nitrophenol in postmitochondrial supernate (S9) fraction of the cells was determined by spectrophotometry. The metabolic activation of HepG2-CYP2E1 cells was assayed by N-nitrosodiethylamine (NDEA)cytotoxicity and micronucleus test.RESULTS: The cloned CYP2E1 cDNA segment was identical to that reported by Umeno et al(GenBank access No.J02843). HepG2-CYP2E1 cells expressed CYP2E1 mRNA and had 4-nitrophenol hydroxylase activity (0.162±0.025nmol.min-1.mg-1 S9 protein), which were undetectable in parent HepG2 cells. HepG2-CYP2E1 cells increased the cytotoxicity and micronucleus rate of NDEA in comparison with those of HepG2 cells.CONCLUSION: The cDNA of human CYP2E1 can be successfully cloned, and a cell line, HepG2-CYP2E1, which can efficiently express mRNA and has CYP2E1 activity, is established. The cell line is useful for testing the cytotoxicity,mutagenicity and metabolism of xenobiotics, which may possibly be activated or metabolized by CYP2E1.

  2. Common Structure of Rare Replication-Deficient E1-Positive Particles in Adenoviral Vector Batches

    Science.gov (United States)

    Murakami, Pete; Havenga, Menzo; Fawaz, Farah; Vogels, Ronald; Marzio, Giuseppe; Pungor, Erno; Files, Jim; Do, Linh; Goudsmit, Jaap; McCaman, Michael

    2004-01-01

    The use of the PER.C6 adenovirus packaging cell line in combination with a designated vector plasmid system, whereby the cell line and vector with E1 deleted have no sequence overlap, eliminates the generation of replication-competent adenovirus during vector production. However, we have found cytopathic effect (CPE)-inducing particles in 2 out of more than 40 large-scale manufacturing lots produced in PER.C6 cells. The CPE inducer was detected at a frequency of 1 event in 7.5 × 1012 vector particles. Despite amplification, it was not readily purified, indicating that the agent itself is replication deficient and requires the parental recombinant adenovirus serotype 5 (rAd5) vector for replication and packaging. Therefore, we designated the agent as a helper-dependent E1-positive region containing viral particle (HDEP). Here, we report the molecular structure of the HDEP genome, revealing an Ad comprised of E1 sequences derived from PER.C6 cells flanked by inverted terminal repeat, packaging signal, and transgene sequences. These sequences form a palindromic structure devoid of E2, E3, E4, and late genes. Since only 5 bp were shared between E1 sequences in the PER.C6 genome and viral vector sequences, the data strongly suggested that insertion of genomic DNA into an adenoviral genome had occurred essentially via nonhomologous recombination. HDEPs have been found in unrelated virus batches and appear to share a common structure that may explain their mechanism of generation. This finding allowed development of an HDEP assay to screen batches of rAd5 produced on the PER.C6 cell line and resulted in detection of seven HDEP agents from four different transgene-virus vector constructs in separate batches of Ad. PMID:15163713

  3. Mouse Spermatocytes Express CYP2E1 and Respond to Acrylamide Exposure

    Science.gov (United States)

    Nixon, Belinda J.; Katen, Aimee L.; Stanger, Simone J.; Schjenken, John E.; Nixon, Brett; Roman, Shaun D.

    2014-01-01

    Metabolism of xenobiotics by cytochrome P450s (encoded by the CYP genes) often leads to bio-activation, producing reactive metabolites that interfere with cellular processes and cause DNA damage. In the testes, DNA damage induced by xenobiotics has been associated with impaired spermatogenesis and adverse effects on reproductive health. We previously reported that chronic exposure to the reproductive toxicant, acrylamide, produced high levels of DNA damage in spermatocytes of Swiss mice. CYP2E1 metabolises acrylamide to glycidamide, which, unlike acrylamide, readily forms adducts with DNA. Thus, to investigate the mechanisms of acrylamide toxicity in mouse male germ cells, we examined the expression of the CYP, CYP2E1, which metabolises acrylamide. Using Q-PCR and immunohistochemistry, we establish that CYP2E1 is expressed in germ cells, in particular in spermatocytes. Additionally, CYP2E1 gene expression was upregulated in these cells following in vitro acrylamide exposure (1 µM, 18 h). Spermatocytes were isolated and treated with 1 µM acrylamide or 0.5 µM glycidamide for 18 hours and the presence of DNA-adducts was investigated using the comet assay, modified to detect DNA-adducts. Both compounds produced significant levels of DNA damage in spermatocytes, with a greater response observed following glycidamide exposure. A modified comet assay indicated that direct adduction of DNA by glycidamide was a major source of DNA damage. Oxidative stress played a small role in eliciting this damage, as a relatively modest effect was found in a comet assay modified to detect oxidative adducts following glycidamide exposure, and glutathione levels remained unchanged following treatment with either compound. Our results indicate that the male germ line has the capacity to respond to xenobiotic exposure by inducing detoxifying enzymes, and the DNA damage elicited by acrylamide in male germ cells is likely due to the formation of glycidamide adducts. PMID:24788432

  4. Repression of insulin gene expression by adenovirus type 5 E1a proteins.

    OpenAIRE

    1987-01-01

    Insulin gene transcription relies on enhancer and promoter elements which are active in pancreatic beta cells. We showed that adenovirus type 5 infection of HIT T-15 cells, a transformed hamster beta cell line, represses insulin gene transcription and mRNA levels. Using expression plasmids transiently introduced into HIT T-15 cells, we showed that adenovirus type 5 E1a transcription regulatory proteins repress insulin enhancer-promoter element activity as assayed with a surrogate xanthine-gua...

  5. Ultrasound associated uptake of chitosan nanoparticles in MC3T3-E1 cells

    Science.gov (United States)

    Wu, Junyi

    Chitosan is a natural linear polysaccharide that has been well known for its applications in drug delivery system due to its unique physicochemical and biological properties. However, challenges still remain for it to become a fully realized therapeutic agent. In this study, we investigated the uptake of chitosan nanoparticles (CNP) under the ultrasound stimulation, using a model cell culture system (MC3T3-E1 mouse pre-osteoblasts). The CNP were fabricated by an ionic gelation method and were lyophilized prior to characterization and delivery to cells. Particle size and zeta potential were measured using Dynamic Light Scattering (DLS); the efficiency of chitosan complexation was measured using the ninhydrin assay. Cytotoxicity was examined by neutral red assay within 48 hours after delivery. The effect of ultrasound (US) on the efficiency of nanoparticle delivery to the MC3T3-E1 cells was examined at 1MHz and at either 1 or 2 W/cm2. Fluorescein isothiocyanate (FITC)-conjugated-CNP were used to visualize the internalized particles within the cytosol. The uptake of FITC-CNP exhibits a dose and time dependent effect, a strong FITC fluorescence was detected at the concentration of 500microg/mL under fluorescence microscope. Ultrasound assisted uptake of FITC-CNP performed a significant positive effect at 2W/cm2 with 60s of ultrasound exposure time. CNP displayed a slightly decrease in cell viability from 25microg/mL to 100microg/mL, while higher concentration of CNP facilitates the proliferation of MC3T3-E1 cells. Less than 10% of reduction in cell viability was observed for US at 1W/cm2 and 2W/cm2 with 30s and 60s of exposure time, which suggest a mild effect of US to MC3T3-E1 cell line.

  6. Calculation of Radiative Corrections to E1 matrix elements in the Neutral Alkalis

    Energy Technology Data Exchange (ETDEWEB)

    Sapirstein, J; Cheng, K T

    2004-09-28

    Radiative corrections to E1 matrix elements for ns-np transitions in the alkali metal atoms lithium through francium are evaluated. They are found to be small for the lighter alkalis but significantly larger for the heavier alkalis, and in the case of cesium much larger than the experimental accuracy. The relation of the matrix element calculation to a recent decay rate calculation for hydrogenic ions is discussed, and application of the method to parity nonconservation in cesium is described.

  7. Migraine: possible role of platelet insensitivity to prostaglandin E1 (PGE1).

    Science.gov (United States)

    Cerneca, F; de Luyk, S; Radillo, O; Simeone, R; Mangiarotti, M

    1993-01-01

    Platelet aggregation inhibition, induced by prostaglandin E1 (PGE1), was evaluated in 38 patients affected by migraine. Our data indicate a complete insensitivity to PGE1 in these subjects. The insensitivity to PGE1 leads to decreased cyclic-AMP (cAMP) levels, determining an imbalance in the inhibitory mechanism. From this observation we can suppose that the decreased affinity of PGE1-receptors, causing decreased cAMP levels, may be involved in pathogenesis of migraine.

  8. E1DS: catalytic site prediction based on 1D signatures of concurrent conservation

    OpenAIRE

    2008-01-01

    Large-scale automatic annotation of protein sequences remains challenging in postgenomics era. E1DS is designed for annotating enzyme sequences based on a repository of 1D signatures. The employed sequence signatures are derived using a novel pattern mining approach that discovers long motifs consisted of several sequential blocks (conserved segments). Each of the sequential blocks is considerably conserved among the protein members of an EC group. Moreover, a signature includes at least thre...

  9. Definition of a major p53 binding site on Ad2E1B58K protein and a possible nuclear localization signal on the Ad12E1B54K protein.

    Science.gov (United States)

    Grand, R J; Parkhill, J; Szestak, T; Rookes, S M; Roberts, S; Gallimore, P H

    1999-01-28

    Previous studies have established that adenovirus 2/5 early region 1B (Ad E1B) 58K protein binds p53 strongly and co-localizes with it to cytoplasmic dense bodies whilst the homologous Ad12E1B54K protein binds only weakly and co-localizes primarily to the nucleus in Ad12E1 transformed cells. We have used these properties of the E1B proteins from different viral serotypes to map the p53 binding site on the Ad2/5 protein. A set of chimaeric genes was constructed containing different proportions of the Ad12 and Ad2E1B DNA. These, together with Ad12E1A and E1B19K DNA, were transfected into baby rat kidney cells and transformed lines isolated. From an examination of the properties of these Ad12/Ad2E1B fusion proteins in co-immunoprecipitation and subcellular localization experiments it has been concluded that the p53 binding site on Ad2E1B58K protein lies between amino acids 216 and 235 and that the homologous region on Ad12E1B54K protein also binds p53. In addition, a unique nuclear localization signal is located on Ad12E1B54K between residues 228 and 239. We suggest that primary structure differences in these regions of the Ad2 and Ad12E1B proteins are responsible for the different subcellular localizations in AdE1 transformants.

  10. CYP1A1 and CYP2E1 polymorphism frequencies in a large Brazilian population

    Directory of Open Access Journals (Sweden)

    Renata dos Santos Coura

    2007-01-01

    Full Text Available The enzymes encoded by the polymorphic genes CYP1A1 and CYP2E1 play an important role in the activation and inactivation of xenobiotics. These enzymes have been associated with xenobiotic-induced diseases, such as cancer, therapeutic failure and adverse effects of drugs. The aim of the present study was to determine the allelic and genotypic frequencies of these polymorphisms in a large, ethnically mixed Brazilian population sample from Rio de Janeiro. Polymorphisms CYP1A1 and CYP2E1 were determined in 870 unrelated individuals by PCR-RFLP analysis in peripheral blood DNA. The observed allelic frequencies were 0.90 for CYP1A1*1A and 0.95 for CYP2E1*1A, in the total sample. The allelic frequency of CYP1A1*2C in "pardos" (0.13 and Brazilian whites (0.11 was higher than in Caucasians (0.05, which may be a result of the Amerindian genetic component, that presents the highest frequency of this allele observed up to now. The genotype distributions for both polymorphisms were in Hardy-Weinberg equilibrium and were statistically different between males and females, and among ethnic groups.

  11. Construction and in vitro Study of an E1B-Defective Adenovirus

    Institute of Scientific and Technical Information of China (English)

    Xue Feng; Joshua Mallam Nock; Zhu Hua-bin; Dong Chang-yuan; Qi Yi-peng

    2004-01-01

    An E1B-defective adenovirus named r1/Ad was constructed by homologous recombination. The construction, selection and propagation of recombinant virus was done in the human embryonic kidney 293 cells (HEK293). The in vitro study demonstrated that the recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as the human glioblastoma tumor cells (U251) and human bladder tumer cells (EJ) but not in the normal cells with functional p53 such as the human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated that the U251 cells were more sensitive to the infection of r1/Ad than that of EJ cells under identical conditions. In this paper, it was found that r1/Ad could be very useful in studying the in vitro selective replication of E1B-defective adenovirus. This may help to determine the safety of using any E1B-defective adenoviruses in cancer gene therapy.

  12. Ubiquitination independent of E1 and E2 enzymes by bacterial effectors

    Energy Technology Data Exchange (ETDEWEB)

    Qiu, Jiazhang; Sheedlo, Michael J.; Yu, Kaiwen; Tan, Yunhao; Nakayasu, Ernesto S.; Das, Chittaranjan; Liu, Xiaoyun; Luo, Zhao-Qing

    2016-04-06

    Signaling by ubiquitination regulates virtually every cellular process in eukaryotes. Covalent attachment of ubiquitin to a substrate is catalyzed by the E1, E2 and E3 three-enzyme cascade 1, which links the C terminus of ubiquitin via an isopeptide bond mostly to the ε-amino group of a lysine of the substrate. Given the essential roles of ubiquitination in the regulation of the immune system, it is not surprising that the ubiquitination network is a common target for diverse infectious agents 2. For example, many bacterial pathogens exploit ubiquitin signaling using virulence factors that function as E3 ligases, deubiquitinases 3 or as enzymes that directly attack ubiquitin 4. The bacterial pathogen Legionella pneumophila utilizes approximately 300 effectors that modulate diverse host processes to create a niche permissive for its replication in phagocytes 5. Here we demonstrate that members of the SidE effector family (SidEs) of L. pneumophila ubiquitinate multiple Rab small GTPases associated with the endoplasmic reticulum (ER). Moreover, we show that these proteins are capable of catalyzing ubiquitination without the need for the E1 and E2 enzymes. The E1/E2-independent ubiquitination catalyzed by these enzymes requires NAD but not ATP and Mg2+. A putative mono ADP-ribosyltransferase (mART) motif critical for the ubiquitination activity is also essential for the role of SidEs in intracellular bacterial replication in a protozoan host. These results establish that ubiquitination can be catalyzed by a single enzyme.

  13. The E1 protein is mandatory for pore formation by Semliki Forest virus spikes.

    Science.gov (United States)

    Dick, M; Barth, B U; Kempf, C

    1996-06-01

    Insect cells (Aedes albopictus, clone C6/36) were infected with various variants of Semliki Forest virus including the wild type using the SFV replicon system. The variants included deletion mutants lacking one of the structural proteins and a mutant with a point mutation in p62 (SQL). The latter mutation results in a failure to process p62 to E2 and E3. After infection of the cells with different variant viruses and subsequent expression of viral proteins in the host cell plasma membrane low pH-induced pore formation was detected by measuring the efflux of a radiolabeled compound. The results of these experiments clearly showed that the E1 protein is mandatory for the acid-induced pore formation. A participation of the 6K or C-protein could be excluded. Furthermore, results obtained with the SQL mutant suggest that dissociation of the E1/E2 heterodimer and subsequent homooligomerization of E1 are required for pore formation.

  14. Effects of 6-Hydroxyflavone on Osteoblast Differentiation in MC3T3-E1 Cells

    Directory of Open Access Journals (Sweden)

    Chien-Hung Lai

    2014-01-01

    Full Text Available Osteoblast differentiation plays an essential role in bone integrity. Isoflavones and some flavonoids are reported to have osteogenic activity and potentially possess the ability to treat osteoporosis. However, limited information concerning the osteogenic characteristics of hydroxyflavones is available. This study investigates the effects of various hydroxyflavones on osteoblast differentiation in MC3T3-E1 cells. The results showed that 6-hydroxyflavone (6-OH-F and 7-hydroxyflavone (7-OH-F stimulated ALP activity. However, baicalein and luteolin inhibited ALP activity and flavone showed no effect. Up to 50 μM of each compound was used for cytotoxic effects study; flavone, 6-OH-F, and 7-OH-F had no cytotoxicity on MC3T3-E1 cells. Moreover, 6-OH-F activated AKT and serine/threonine kinases (also known as protein kinase B or PKB, extracellular signal-regulated kinases (ERK 1/2, and the c-Jun N-terminal kinase (JNK signaling pathways. On the other hand, 7-OH-F promoted osteoblast differentiation mainly by activating ERK 1/ 2 signaling pathways. Finally, after 5 weeks of 6-OH-F induction, MC3T3-E1 cells showed a significant increase in the calcein staining intensity relative to merely visible mineralization observed in cells cultured in the osteogenic medium only. These results suggested that 6-OH-F could activate AKT, ERK 1/2, and JNK signaling pathways to effectively promote osteoblastic differentiation.

  15. In vivo and in vitro characterization of CYP2E1 activity in Japanese and Caucasians.

    Science.gov (United States)

    Kim, R B; Yamazaki, H; Chiba, K; O'Shea, D; Mimura, M; Guengerich, F P; Ishizaki, T; Shimada, T; Wilkinson, G R

    1996-10-01

    Chlorzoxazone's disposition after oral administration was determined in 20 young healthy Caucasian men and a similar group of Japanese men. The drug's plasma concentrations were significantly higher and its rate of elimination slower in Japanese compared to Caucasian men. Accordingly, chlorzoxazone's oral clearance was smaller (40%) in Japanese men and a similar difference (30%) was still apparent after normalizing for body weight (3.74 +/- 1.23 versus 5.05 +/- 1.41 ml.min-1.kg-1, P relationships were found between measures of CYP2E1 activity, both in vivo and in vitro, and genomic polymorphisms in the CYP2E1 gene identified by Rsal/Pstl and Dral restriction fragment length polymorphisms. Collectively, these data show an interracial difference in CYP2E1 activity. Because this enzyme is importantly involved in the activation of environmental procarcinogens, such a difference may account, in part, for the lower rate of some cancers, e.g., lung cancer, in Japanese compared to Caucasians men.

  16. Electric dipole strength distribution below the E1 giant resonance in N = 82 nuclei

    Science.gov (United States)

    Guliyev, Ekber; Kuliev, Ali; Guner, Mehmet

    2010-12-01

    In this study quasiparticle random-phase approximation with the translational invariant Hamiltonian using deformed mean field potential has been conducted to describe electric dipole excitations in 136Xe, 138Ba, 140Ce, 142Nd, 144Sm and 146Gd isotones. The distribution of the calculated E1 strength shows a resonance like structure at energies between 6-8 MeV exhausting up to 1% of the isovector electric dipole Energy Weighted Sum Rule and in some aspects nicely confirms the experimental data. It has been shown that the main part of E1 strength, observed below the threshold in these nuclei may be interpreted as main fragments of the Pygmy Dipole resonance. The agreement between calculated mean excitation energies as well as summed B(E1) value of the 1- excitations and the available experimental data is quite good. The calculations indicate the presence of a few prominent positive parity 1+ States in heavy N = 82 isotones in the energy interval 6-8 MeV which shows not all dipole excitations were of electric character in this energy range.

  17. Instanton induced Yukawa couplings from distant E3 and E(-1) instantons

    Energy Technology Data Exchange (ETDEWEB)

    Goodsell, Mark D. [Sorbonne Universités, UPMC Univ Paris 06, UMR 7589, LPTHE,F-75005, Paris (France); CNRS, UMR 7589, LPTHE,F-75005, Paris (France); Witkowski, Lukas T. [Institute for Theoretical Physics, University of Heidelberg,Philosophenweg 19, 69120 Heidelberg (Germany)

    2016-01-07

    We calculate non-perturbative contributions to Yukawa couplings on D3-branes at orbifold singularities due to E3 and fractional E(-1) instantons which do not intersect the visible sector branes. While distant E3 instantons on bulk cycles typically contribute to Yukawa couplings, we find that distant fractional E(-1) can also give rise to new Yukawa couplings. However, fractional E(-1) instantons only induce Yukawa couplings if they are located at a singularity which shares a collapsed homologous two-cycle with the singularity supporting the visible sector. The non-perturbative contributions to Yukawa couplings exhibit a different flavour structure than the tree-level Yukawa couplings and, as a result, they can be sources of flavour violation. This is particularly relevant for schemes of moduli stabilisation which rely on superpotential contributions from E3 instantons, such as KKLT or the Large Volume Scenario. As a byproduct of our analysis, we shed some new light on the properties of annulus diagrams with matter field insertions in stringy instanton calculus.

  18. Evidence for selective regulation of the phosphorylation of myocyte proteins by isoproterenol and prostaglandin E1.

    Science.gov (United States)

    Hayes, J S; Bowling, N; King, K L; Boder, G B

    1982-01-12

    Both isoproterenol and prostaglandin E1 increased the activation state of cyclic AMP-dependent protein kinase in cultured myocytes; however, only isoproterenol enhanced phosphorylase activity and contractile state. Following the incubation of intact myocytes with 32PO3-(4), 32 phosphoproteins were resolved from total cellular proteins by electrophoresis in sodium dodecyl sulfate polyacrylamide gels followed by autoradiography. Isoproterenol stimulated 32PO3-(4) incorporation into 16 proteins, including 2 phosphoproteins not observed under control conditions. By contrast, prostaglandin E1 neither caused a measurable change in the protein phosphorylation pattern nor interfered with isoproterenol's capacity to do so. Isoproterenol stimulated myocyte protein phosphorylation in either the presence or absence of extracellular Ca2+. The results suggest that the regulation of protein phosphorylation following adenylate cyclase stimulation is: (1) an agonist-specific process and not due solely to a random accumulation of intracellular cycle AMP and activation of protein kinase; (2) the Ca2+ mobilization component of beta-receptor activation does not account for the paradoxical effects of isoproterenol and prostaglandin E1; (3) activation of cyclic AMP-dependent protein kinase does not always result in an enhancement of protein phosphorylation.

  19. Ovarian Hormone Estrone Glucuronide (E1G) quantification-impedimetric electrochemical spectroscopy approach

    KAUST Repository

    Zia, Asif I.

    2013-12-01

    A study was conducted on detection and concentration measurement of estrone glucuronide (E1G), an important metabolite of the ovarian hormone estradiol, by using Electrochemical Impedance Spectroscopy (EIS) technique. A miniature planar Inter-digital (ID) capacitive sensor fabricated on single crystal silicon substrate with sputtered gold electrodes coupled with EIS was used to measure conductivity, permeability and dielectric properties of the said hormone metabolite. A thin film of Silicon Nitride (50 um) was coated on the sensor as passivation layer to avoid Faradic currents through the sensor. Impedance spectrums were obtained with various concentrations of E1G in buffer solution by exposing the samples to electrical perturbations at certain frequency range. Relationship of sample conductance with E1G concentration was studied on basis Randle\\'s equivalent circuit model and results were analyzed to deduce Constant Phase Equivalent (CPE) Circuit model in order to evaluate the double layer capacitance produced at the solution-electrode interface due to kinetic processes taking place in the electrochemical cell. The sensitivity of the sensor was evaluated against concentration. The result analysis confirmed that fabricated ID sensor together with EIS can provide a rapid and successful low cost sensing system which can help a lay user to determine peak time for feminine reproductive fertility at home without submitting samples for an expensive and time consuming laboratory test. © 2013 IEEE.

  20. Identification of Interactions in the E1E2 Heterodimer of Hepatitis C Virus Important for Cell Entry*

    Science.gov (United States)

    Maurin, Guillemette; Fresquet, Judith; Granio, Ophélia; Wychowski, Czeslaw; Cosset, François-Loïc; Lavillette, Dimitri

    2011-01-01

    Several conserved domains critical for E1E2 assembly and hepatitis C virus entry have been identified in E1 and E2 envelope glycoproteins. However, the role of less conserved domains involved in cross-talk between either glycoprotein must be defined to fully understand how E1E2 undergoes conformational changes during cell entry. To characterize such domains and to identify their functional partners, we analyzed a set of intergenotypic E1E2 heterodimers derived from E1 and E2 of different genotypes. The infectivity of virions indicated that Con1 E1 did not form functional heterodimers when associated with E2 from H77. Biochemical analyses demonstrated that the reduced infectivity was not related to alteration of conformation and incorporation of Con1 E1/H77 E2 heterodimers but rather to cell entry defects. Thus, we generated chimeric E1E2 glycoproteins by exchanging different domains of each protein in order to restore functional heterodimers. We found that both the ectodomain and transmembrane domain of E1 influenced infectivity. Site-directed mutagenesis highlighted the role of amino acids 359, 373, and 375 in transmembrane domain in entry. In addition, we identified one domain involved in entry within the N-terminal part of E1, and we isolated a motif at position 219 that is critical for H77 function. Interestingly, using additional chimeric E1E2 complexes harboring substitutions in this motif, we found that the transmembrane domain of E1 acts as a partner of this motif. Therefore, we characterized domains of E1 and E2 that have co-evolved inside a given genotype to optimize their interactions and allow efficient entry. PMID:21555519

  1. The application of the instantaneous states reduction to the orbital monitoring of pivotal arcs of the Chang’E-1 satellite

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    In the Chinese lunar exploration project,the Chang’E-1 (CE-1) satellite was jointly monitored by the United S-band range and Doppler and the VLBI technique. A real-time reduction of the tracking data is realized to deduce the time series of the instantaneous state vectors (ISV) (position and velocity vec-tors) of the CE-1 satellite,and is applied to the orbital monitoring of pivotal arcs. This paper introduces this real-time data reduction method and its application to the orbital monitoring of pivotal arcs of the CE-1 satellite in order to serve as a source of criticism and reference.

  2. (E-1-(2-Aminophenyl-3-(benzo[d][1,3]dioxol-5-ylprop-2-en-1-one

    Directory of Open Access Journals (Sweden)

    Rodrigo Abonia

    2016-12-01

    Full Text Available The title chalcone (E-1-(2-aminophenyl-3-(benzo[d][1,3]dioxol-5-ylprop-2-en-1-one was obtained in 76% yield from a NaOH catalyzed Claisen–Schmidt condensation reaction between o-aminoacetophenone and piperonal. This product will be used as a key precursor for the development of an alternative route for the total synthesis of the alkaloid Graveoline. Single crystals of the title compound suitable for X-ray diffraction were grown via slow evaporation in ethanol at room temperature. A complete crystallographic study was performed in depth to unequivocally confirm its structure. The crystal structure of the title o-aminochalcone, C16H13NO3, shows two molecules per asymmetric unit (Z = 4 and adopts an E configuration about the C=C double bond. In the title compound, the mean plane of the non-H atoms of the central chalcone fragment C—C(O—C—C—C is as follow: [root-mean-square (r.m.s. deviation = 0.0210 Å for A–B and 0.0493 for C–D molecules]. In the crystal, molecules are linked by N–H...O and C–H...O, hydrogen bonds forming S(6, R22(6 and edge-fused R44(24rings along with C(18 chains running parallel to (110.

  3. Effects of ethanol on CYP2E1 levels and related oxidative stress using a standard balanced diet.

    Science.gov (United States)

    Azzalis, Ligia A; Fonseca, Fernando L A; Simon, Karin A; Schindler, Fernanda; Giavarotti, Leandro; Monteiro, Hugo P; Videla, Luis A; Junqueira, Virgínia B C

    2012-07-01

    Expression of cytochrome P4502E1 (CYP2E1) is very much influenced by nutritional factors, especially carbohydrate consumption, and various results concerning the expression of CYP2E1 were obtained with a low-carbohydrate diet. This study describes the effects of ethanol treatment on CYP2E1 levels and its relationship with oxidative stress using a balanced standard diet to avoid low or high carbohydrate consumption. Rats were fed for 1, 2, 3, or 4 weeks a commercial diet plus an ethanol-sucrose solution. The results have shown that ethanol administration was associated with CYP2E1 induction and stabilization without related oxidative stress. Our findings suggest that experimental models with a low-carbohydrate/high-fat diet produce some undesirable CYP2E1 changes that are not present when a balanced standard diet is given.

  4. Structure of Pyrazole Derivatives Impact their Affinity, Stoichiometry, and Cooperative Interactions for CYP2E1 Complexes

    OpenAIRE

    2013-01-01

    CYP2E1 plays a critical role in detoxication and carcinogenic activation of drugs, pollutants, and dietary compounds; however, these metabolic processes can involve poorly characterized cooperative interactions that compromise the ability to understand and predict CYP2E1 metabolism. Herein, we employed an array of ten azoles with an emphasis on pyrazoles to establish the selectivity of catalytic and cooperative CYP2E1 sites through binding and catalytic studies. Spectral binding studies for m...

  5. Evidence for the two-body nature of the E1 transition operator in the sdf-interacting boson model

    NARCIS (Netherlands)

    Barfield, A. F.; von Brentano, P.; Dewald, A.; Zell, K. O.; Zamfir, N. V.; Bucurescu, D.; Ivaşcu, M.; Scholten, O.

    1989-01-01

    Two new absolute transition rates are reported for the nucleus144Sm following an ( α, α') Coulomb excitation study. They are B(E3; 3-→ 0+)=(38±3) W.u. and B(E1;3- → 2+)=(2.8±0.4)×10-3 W.u. This large E1 matrix element, along with the previously known B(E1; 1- →+) value support the interpretation of

  6. Structure of Pyrazole Derivatives Impact their Affinity, Stoichiometry, and Cooperative Interactions for CYP2E1 Complexes

    OpenAIRE

    Hartman, Jessica H.; Bradley, Amber M.; Laddusaw, Ryan M.; Perry, Martin D.; Miller, Grover P.

    2013-01-01

    CYP2E1 plays a critical role in detoxication and carcinogenic activation of drugs, pollutants, and dietary compounds; however, these metabolic processes can involve poorly characterized cooperative interactions that compromise the ability to understand and predict CYP2E1 metabolism. Herein, we employed an array of ten azoles with an emphasis on pyrazoles to establish the selectivity of catalytic and cooperative CYP2E1 sites through binding and catalytic studies. Spectral binding studies for m...

  7. ON SPACELIKE BIHARMONIC SLANT HELICES ACCORDING TO BISHOP FRAME IN THE LORENTZIAN GROUP OF RIGID MOTIONS E(1,1

    Directory of Open Access Journals (Sweden)

    Talat Körpınar

    2012-01-01

    Full Text Available In this paper, we study spacelike biharmonic slant helices according to Bishop frame in the Lorentzian group of rigid motions E(1,1. We characterize the spacelike biharmonic slant helices in terms of their curvatures in the Lorentzian group of rigid motions E(1,1. Finally, we obtain parametric equations of spacelike biharmonic slant helices according to Bishop frame in the Lorentzian group of rigid motions E(1,1.

  8. Ubiquitination independent of E1 and E2 enzymes by bacterial effectors.

    Science.gov (United States)

    Qiu, Jiazhang; Sheedlo, Michael J; Yu, Kaiwen; Tan, Yunhao; Nakayasu, Ernesto S; Das, Chittaranjan; Liu, Xiaoyun; Luo, Zhao-Qing

    2016-05-01

    Signalling by ubiquitination regulates virtually every cellular process in eukaryotes. Covalent attachment of ubiquitin to a substrate is catalysed by the E1, E2 and E3 three-enzyme cascade, which links the carboxy terminus of ubiquitin to the ε-amino group of, in most cases, a lysine of the substrate via an isopeptide bond. Given the essential roles of ubiquitination in the regulation of the immune system, it is not surprising that the ubiquitination network is a common target for diverse infectious agents. For example, many bacterial pathogens exploit ubiquitin signalling using virulence factors that function as E3 ligases, deubiquitinases or as enzymes that directly attack ubiquitin. The bacterial pathogen Legionella pneumophila utilizes approximately 300 effectors that modulate diverse host processes to create a permissive niche for its replication in phagocytes. Here we demonstrate that members of the SidE effector family of L. pneumophila ubiquitinate multiple Rab small GTPases associated with the endoplasmic reticulum. Moreover, we show that these proteins are capable of catalysing ubiquitination without the need for the E1 and E2 enzymes. A putative mono-ADP-ribosyltransferase motif critical for the ubiquitination activity is also essential for the role of the SidE family in intracellular bacterial replication in a protozoan host. The E1/E2-independent ubiquitination catalysed by these enzymes is energized by nicotinamide adenine dinucleotide, which activates ubiquitin by the formation of ADP-ribosylated ubiquitin. These results establish that ubiquitination can be catalysed by a single enzyme, the activity of which does not require ATP.

  9. Cooperativity in CYP2E1 metabolism of acetaminophen and styrene mixtures.

    Science.gov (United States)

    Hartman, Jessica H; Letzig, Lynda G; Roberts, Dean W; James, Laura P; Fifer, E Kim; Miller, Grover P

    2015-10-01

    Risk assessment for exposure to mixtures of drugs and pollutants relies heavily on in vitro characterization of their bioactivation and/or metabolism individually and extrapolation to mixtures assuming no interaction. Herein, we demonstrated that in vitro CYP2E1 metabolic activation of acetaminophen and styrene mixtures could not be explained through the Michaelis-Menten mechanism or any models relying on that premise. As a baseline for mixture studies with styrene, steady-state analysis of acetaminophen oxidation revealed a biphasic kinetic profile that was best described by negative cooperativity (Hill coefficient=0.72). The best-fit mechanism for this relationship involved two binding sites with differing affinities (Ks=830μM and Kss=32mM). Introduction of styrene inhibited that reaction less than predicted by simple competition and thus provided evidence for a cooperative mechanism within the mixture. Likewise, acetaminophen acted through a mixed-type inhibition mechanism to impact styrene epoxidation. In this case, acetaminophen competed with styrene for CYP2E1 (Ki=830μM and Ksi=180μM for catalytic and effector sites, respectively) and resulted in cooperative impacts on binding and catalysis. Based on modeling of in vivo clearance, cooperative interactions between acetaminophen and styrene resulted in profoundly increased styrene activation at low styrene exposure levels and therapeutic acetaminophen levels. Current Michaelis-Menten based toxicological models for mixtures such as styrene and acetaminophen would fail to detect this concentration-dependent relationship. Hence, future studies must assess the role of alternate CYP2E1 mechanisms in bioactivation of compounds to improve the accuracy of interpretations and predictions of toxicity.

  10. Laser altimetry data of Chang’E-1 and the global lunar DEM model

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The Laser AltiMeter (LAM), as one of the main payloads of Chang’E-1 probe, is used to measure the topography of the lunar surface. It performed the first measurement at 02:22 on November 28th, 2007. Up to December 4th 2008, the total number of measurements was approximately 9.12 million, covering the whole surface of the Moon. Using the LAM data, we constructed a global lunar Digtal Elevation Model (DEM) with 3 km spatial resolution. The model shows pronounced morphological characteristics, legible and vivid details of the lunar surface. The plane positioning accuracy of the DEM is 445 m (1σ), and the vertical accuracy is 60 m (1σ). From this DEM model, we measured the full range of the altitude difference on the lunar sur-face, which is about 19.807 km. The highest point is 10.629 km high, on a peak between crater Korolev and crater Dirichlet-Jackson at (158.656°W, 5.441°N) and the lowest point is -9.178 km in height, inside crater Antoniadi (172.413°W, 70.368°S) in the South Pole-Aitken Basin. By comparison, the DEM model of Chang’E-1 is better than the USA ULCN2005 in accuracy and resolution and is probably identical to the DEM of Japan SELENE, but the DEM of Chang’E-1 reveals a new lowest point, clearly lower than that of SELENE.

  11. Lysophosphatidic acid induces chemotaxis in MC3T3-E1 osteoblastic cells

    Energy Technology Data Exchange (ETDEWEB)

    Masiello, Lisa M.; Fotos, Joseph S.; Galileo, Deni S.; Karin, Norm J.

    2006-07-01

    Lysophosphatidic acid (LPA) is a bioactive lipid that has pleiotropic effects on a variety of cell types and enhances the migration of endothelial and cancer cells, but it is not known if this lipid can alter osteoblast motility. We performed transwell migration assays using MC3T3-E1 osteoblastic cells and found LPA to be a potent chemotactic agent. Quantitative time-lapse video analysis of osteoblast migration after wounds were introduced into cell monolayers indicated that LPA stimulated both migration velocity and the average migration distance per cell. LPA also elicited substantial changes in cell shape and actin cytoskeletal structure; lipid-treated cells contained fewer stress fibers and displayed long membrane processes that were enriched in F-actin. Quantitative RT-PCR analysis showed that MC3T3-E1 cells express all four known LPA-specific G protein-coupled receptors (LPA1-LPA4) with a relative mRNA abundance of LPA1 > LPA4 > LPA2 >> LPA3. LPA-induced changes in osteoblast motility and morphology were antagonized by both pertussis toxin and Ki16425, a subtype-specific blocker of LPA1 and LPA3 receptor function. Cell migration in many cell types is linked to changes in intracellular Ca2+. Ki16425 also inhibited LPA-induced Ca2+ signaling in a dose-dependent manner, suggesting a link between LPA-induced Ca2+ transients and osteoblast chemotaxis. Our data show that LPA stimulates MC3T3-E1 osteoblast motility via a mechanism that is linked primarily to the G protein-coupled receptor LPA1.

  12. Occupational Survey Report. Ground Radio Communications, AFSC 2E1X3

    Science.gov (United States)

    2003-10-01

    analyzed the data, and wrote the final report. Mr. Tyrone Hill provided computer-programming support, and Ms. Dolores Navarro and Ms. Sherry Evans...AFSC 2E1X3 members selected military-related education and training opportunities and medical/ dental care for AD members as their second and third...TRAINING OPPORTUNITIES 52 2.52 55 2.49 50 2.34 MEDICAL/ DENTAL CARE FOR AD MEMBER 60 2.57 55 2.56 53 2.48 MEDICAL/ DENTAL CARE FOR FAMILY MEMBERS 48

  13. Cytochrome P450 2E1 genetic polymorphism and gastric cancer in Changle, Fujian Province

    Institute of Scientific and Technical Information of China (English)

    Lin Cai; Shun-Zhang Yu; Zuo-Feng Zhang

    2001-01-01

    AIM: Genetic polymorphism in enzymes of carcinogen metabolism has been found to have the influence on the susceptibility to cancer. Cytochrome P450 2E1 ( CYP2 E1) is considered to play an important role in the metabolic activation of procarcinogens such as N-nitroscoamines and Iow molecular weight organic compounds. The purpose of this study is to determine whether CYP450 2Elpolymorphisms are associated with risk s of gastric cancer. METHODS: We conducted a population based case-control study in Changle county, Fujian Province, a high-risk region of gastric cancer in China. Ninety-one incident gastric cancer patients and ninety-four healthy controls were included in our study. Datas including dsmographic characteristcs, diet intake, and alcohol and tobacco consumption of indivduals in our study were completed by a standardized questionnaire. PCR-RFLP revealed three genotypes: heterozygote (C1/C2) and two homozygotes (C1/C1 and C2/C2) in CYP2E1. RESULTS: The frequency of variant genotypes (C1/C2 and C2/C2) in gastric cancer cases and controls was 36.3% and 24.5%, respectively. The rare homozygous C2/C2 genotype was found in 6 indivduals in gastric cancer group(6.6%),whereas there was only one in the control group (1.1%).However, there was no statistically significan difference between the two groups (two-tailed Fisher′s exact test, P =0.066). Indivduals in gastric cancer group were more likely to carry genotype C1/C2 (odds ratio, OR = 1.50) and C2/C2(OR = 7.34) than indivduals in control group (X2 = 4.597, for trend P=0.032). The frequencies of genofypes with the C2allele ( C1/C2 and C2/C2 genotypes) were compared with those of genotypes without C2 allele ( C1/C1 genotype )among indivduals in gastric cancer group and control group according to the pattern of gastric cancer risk factors. The results show that indivduals who exposed to these gastric cancer risk factors and carry the C2 allele seemed to have a higher risk of developing gastric cancer. CONCLUSION

  14. Analysen zur Funktion des E1B-55K-Proteins von Adenovirus Typ 5 im lytischen Replikationszyklus

    OpenAIRE

    Kindsmüller, Kathrin

    2007-01-01

    Das E1B-55K-Protein von Adenovirus Typ 5 ist ein multifunktionelles Phosphoprotein, das eine zentrale Rolle im produktiven Replikationszyklus von Ad5 einnimmt. Die lytischen Aktivitäten des E1B-Proteins werden zumindest teilweise im Komplex mit dem viralen Protein E4orf6 vermittelt. Soweit bekannt fördert der E1B-55K/E4orf6-Komplex (E1B/E4-Komplex) den nukleozytoplasmatischen Transport viraler mRNAs und fördert somit die Synthese viraler Kapsidproteine bzw. die Produktion von Nachkommenviren....

  15. De-repression of RaRF-mediated RAR repression by adenovirus E1A in the nucleolus.

    Science.gov (United States)

    Um, Soo-Jong; Youn, Hye Sook; Kim, Eun-Joo

    2014-02-21

    Transcriptional activity of the retinoic acid receptor (RAR) is regulated by diverse binding partners, including classical corepressors and coactivators, in response to its ligand retinoic acid (RA). Recently, we identified a novel corepressor of RAR called the retinoic acid resistance factor (RaRF) (manuscript submitted). Here, we report how adenovirus E1A stimulates RAR activity by associating with RaRF. Based on immunoprecipitation (IP) assays, E1A interacts with RaRF through the conserved region 2 (CR2), which is also responsible for pRb binding. The first coiled-coil domain of RaRF was sufficient for this interaction. An in vitro glutathione-S-transferase (GST) pull-down assay was used to confirm the direct interaction between E1A and RaRF. Further fluorescence microscopy indicated that E1A and RaRF were located in the nucleoplasm and nucleolus, respectively. However, RaRF overexpression promoted nucleolar translocation of E1A from the nucleoplasm. Both the RA-dependent interaction of RAR with RaRF and RAR translocation to the nucleolus were disrupted by E1A. RaRF-mediated RAR repression was impaired by wild-type E1A, but not by the RaRF binding-defective E1A mutant. Taken together, our data suggest that E1A is sequestered to the nucleolus by RaRF through a specific interaction, thereby leaving RAR in the nucleoplasm for transcriptional activation.

  16. Interaction of Adenovirus E1A with the HHV8 Promoter of Latent Genes: E1A Proteins are Able to Activate the HHV-8 LANAp in MV3 Reporter Cells.

    Science.gov (United States)

    Koehler-Hansner, Karin; Flore, Ornella; Opalka, Bertram; Hengge, Ulrich R

    2008-01-01

    Human herpesvirus 8 (HHV-8) is associated with Kaposi's sarcoma, body cavity-based lymphoma, and Castleman's disease. Adenoviral (Ad) E1A proteins regulate the activity of cellular and viral promoters/enhancers and transcription factors and can suppress tumorigenicity of human cancers. As (i) HHV-8 and Ad may co-exist in immunocompromised patients and (ii) E1A might be considered as therapeutic transgene for HHV-8-associated neoplasms we investigated whether the promoter of the latency-associated nuclear antigen (LANAp) controlling expression of vCyclin, vFLIP, and LANA proteins required for latent type infection is regulated by E1A. Transfection experiments in MV3 melanoma cells revealed activation of the LANAp by Ad5 E1A constructs containing an intact N terminus (aa 1-119). In particular, an Ad12 E1A mutant, Spm2, lacking six consecutive alanine residues in the "spacer" region activated the HHV-8 promoter about 15-fold compared to vector controls. In summary, we report the activation of the LANAp by E1A as a novel interaction of E1A with a viral promoter. These data may have relevance for the management of viral infections in immunocompromised patients. A role for E1A as a therapeutic in this context remains to be defined.

  17. Behavior of MC3T3-E1 Osteoblast Cultured on Chitosan Modified with Polyvinylpyrrolidone

    Institute of Scientific and Technical Information of China (English)

    XI Jing; GAO Yuan; KONG Lijun; GONG Yandao; ZHAO Nanming; ZHANG Xiufang

    2005-01-01

    The physical and chemical properties of four kinds of modified chitosan materials made by blending chitosan with polyvinylpyrrolidone (PVP) were investigated. All four of these modified chitosan materials were hydrophilic with water contact angles ranging from 59° to 69°. Fourier transform-infrared spectra of the modified materials showed a new band at 1288 cm-1, implying formation of a surface physical interpenetrating network structure. Enzyme linked immunosorbent assay results indicated that much less fibronectin was adsorbed on the modified materials than on only chitosan. The viability of MC3T3-E1 osteoblasts cultured on the materials was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl- 2H-tetrazolium bromide assay. The results show that adding PVP10000 into the chitosan promotes adhesion of MC3T3-E1 osteoblasts on the modified materials, but has no effect on cell growth and proliferation; while adding PVP40000 reduces cell adhesion, growth, and proliferation. The results suggest that the increased hydrophilicity of the material surface does not always improve its biocompatibility, which will influence the selection and design of biomaterials.

  18. Lunar topographic model CLTM-s01 from Chang'E-1 laser altimeter

    Institute of Scientific and Technical Information of China (English)

    PING JinSong; HUANG Qian; YAN JianGuo; CAO JianFeng; TANG GeShi; SHU Rong

    2009-01-01

    More than 3 million range measurements from the Chang'E-1 Laser Altimeter have been used to pro-duce a global topographic model of the Moon with improved accuracy. Our topographic model, a 360th degree and order spherical harmonic expansion of the lunar radii, is designated as Chang'E-1 Lunar Topography Model s01 (CLTM-s01). This topographic field, referenced to a mean radius of 1738 km, has an absolute vertical accuracy of approximately 31 m and a spatial resolution of 0.25° (~7.5 km). This new lunar topographic model has greatly improved previous models in spatial coverage, accuracy and spatial resolution, and also shows the polar regions with the altimeter results for the first time. From CLTM-s01, the mean, equatorial, and polar radii of the Moon are 1737103, 1737646, and 1735843 m, re-spectively. In the lunar-fixed coordinate system, this model shows a COM/COF offset to be (-1.777, -0.730, 0.237) km along the x, y, and z directions, respectively. All the basic lunar shape parameters derived from CLTM-s01 are in agreement with the results of Clementine GLTM2, but CLTM-s01 offers higher accuracy and reliability due to its better global samplings.

  19. Lunar topographic model CLTM-s01 from Chang’E-1 laser altimeter

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    More than 3 million range measurements from the Chang’E-1 Laser Altimeter have been used to produce a global topographic model of the Moon with improved accuracy. Our topographic model, a 360th degree and order spherical harmonic expansion of the lunar radii, is designated as Chang’E-1 Lunar Topography Model s01 (CLTM-s01). This topographic field, referenced to a mean radius of 1738 km, has an absolute vertical accuracy of approximately 31 m and a spatial resolution of 0.25o (~7.5 km). This new lunar topographic model has greatly improved previous models in spatial coverage, accuracy and spatial resolution, and also shows the polar regions with the altimeter results for the first time. From CLTM-s01, the mean, equatorial, and polar radii of the Moon are 1737103, 1737646, and 1735843 m, respectively. In the lunar-fixed coordinate system, this model shows a COM/COF offset to be (-1.777, -0.730, 0.237) km along the x, y, and z directions, respectively. All the basic lunar shape parameters derived from CLTM-s01 are in agreement with the results of Clementine GLTM2, but CLTM-s01 offers higher accuracy and reliability due to its better global samplings.

  20. Effects of salvia miltiorrhiza bunge (SMB) on MC3T3-E1 cells.

    Science.gov (United States)

    Ding, Y; Soma, S; Takano-Yamamoto, T; Matsumoto, S; Sakuda, M

    1995-12-01

    In previous research, we found that the Salvia Miltiorrhiza Bunge (SMB), a traditional Chinese medicinal herb, accelerates orthodontic tooth movement. In the present study, to characterize the actions of SMB on bone remodeling, we investigated the effect of SMB on DNA synthesis and alkaline phosphatase activity of murine osteoblast like cell-clone, MC3T3-E1 cells in vitro. Treatment of the cells with SMB for 72 hours caused significant increase in ALPase activity. It was found that SMB increased ALPase activity in a dose-dependent manner, and up to maximum at the concentration of 5.0 mg/ml. At that concentration, ALPase activity was about 135% greater than that of control. SMB at 5.0 mg/ml significantly stimulated ALPase activity of the cells in multilayer on day 8 or calcification stage on day 16, but inhibited it in the sparse stage on day 2 or subconfluency stage on day 4. SMB had no effect on DNA synthesis in any stage of culture. ALPase activity significantly increased at 48 hours and was up to 300% greater than that of control activity at 96 hours. These findings suggest that SMB directly stimulates ALPase activity of MC3T3-E1 cells in multilayer stage or calcification stage without any effect on proliferation. Locally administrated SMB may affect the differentiation of osteoblasts in vivo.

  1. The effect of prostaglandin E1 on recovery of early renal graft functions after transplantation

    Institute of Scientific and Technical Information of China (English)

    Song Huanjin; Xue Wujun; Tian Xiaohui; Li Yang; Ding Chenguang; Ding Xiaoming; Feng Xinshun; Jin Zhankui

    2008-01-01

    Objective To investigate the effect of prostaglandin E: (PGE1) on recovery of early renal graft functions after transplantation. Methods One hundred and seven patients after renal transplantation were allocated in the treated group, and treated by conventional treatment with injection of 10 μg prostaglandin E1 additionally twice a day for 14 days. And eighty-eight patients who received conventional treatment alone after renal transplantation at the corresponding period were allocated in the control group. Indexes of the two groups, including incidence of delayed graft function and acute rejection reaction, volume of urine, serum certaintie (SCr), endogenous certainties clearance rate (CCr), the blood flow resistance in graft as well as blood viscosity (BV), and platelet aggregation rate (PAR), were determined. Results The urinary volume and endogenous certainties clearance rate of the treated group were significantly higher, but the level of SCr, incidence of renal function recovery retardation, BV, PAR and blood flow resistance in graft were significantly lower than these of the control group (P0.05). Conclusion Prostaglandin E1 can improve blood microcirculation and decrease the incidence of renal function recovery retardation. These effects are helpful for recovery of renal function after renal transplantation.

  2. [Genetic polymorphism of cytochrome P450 2E1 and the risk of nasopharyngeal carcinoma].

    Science.gov (United States)

    Ben Chaaben, Arij; Abaza, Hajer; Douik, Hayet; Chaouch, Leila; Ayari, Fayza; Ouni, Nesrine; Mamoghli, Tasnim; Ben Guezella, Dorra; Mejri, Rachida; Harzallah, Latifa; Guemira, Fethi

    2015-12-01

    Cytochrome P450 2E1 (CYP2E1) is a detoxifying enzyme that belongs to the phase I metabolism of xenobiotics. This enzyme is encoded by a highly polymorphic gene whose common polymorphism corresponds to the substitution of cytosine (C) and thymine (T) at position -1019 (rs2031920). This polymorphism has been identified in several cancers including nasopharyngeal cancer (NPC). The study involved 124 patients with nasopharyngeal carcinoma, compared with 166 healthy controls. The presence or absence of the polymorphism is determined by PCR-RFLP. The frequency comparison between the two groups is determined by the χ(2) test. The analysis of our results showed a significant difference between the two groups regarding the mutant genotype (C2/C2) (5% vs. 0.5%, P=0.04) and has a risk factor for NPC in Tunisia (OR=8.39; CI 95% [0.99-388.1]). Also, the C2 allele was significantly associated with the group of patients than the control group (6% vs. 2%, P=0.016) and increased three times the risk of NPC in Tunisia (OR=2.99, CI 95% [1.12-8.79]). Our results confirm the results reported in other populations and emphasize the importance of the involvement of this gene in the development of detoxification of the NPC, which seems more and more strongly associated with environmental factors.

  3. E1 resonances in neutron-rich nuclei within the phonon damping model

    Indian Academy of Sciences (India)

    Nguyen Dinh Dang; Voung Kim Au; Toshio Suzuki; Akito Arima

    2001-08-01

    The quasiparticle representation of the phonon damping model (PDM) is developed to include the superfluid pairing correlations microscopically. The formalism is applied to calculate the photoabsorption and the electromagnetic (EM) differential cross sections of E1 excitations in neutron-rich oxygen and calcium isotopes. The calculated photoabsorption cross sections agree reasonably well with the available data for 16,18O and 40,48Ca. The results of calculations show that the change of the fraction of the E1 integrated strength in the region of pygmy dipole resonance (PDR) as a function of mass number with increasing neutron number is in agreement with the recent experimental data, and does not follow the prediction by the simple cluster model. The EM differential cross sections obtained within PDM in this work show prominent PDR peaks below 15 MeV for 20,22O in agreement with the recent experimental observation. It is also shown that, using low-energy RI beams at around 50-60 MeV/nucleon, one can observe clean and even enhanced PDR peaks without the admixture with the GDR in the EM differential cross sections of neutron-rich nuclei.

  4. Comparison of chlorzoxazone one-sample methods to estimate CYP2E1 activity in humans

    DEFF Research Database (Denmark)

    Kramer, Iza; Dalhoff, Kim; Clemmesen, Jens O

    2003-01-01

    OBJECTIVE: Comparison of a one-sample with a multi-sample method (the metabolic fractional clearance) to estimate CYP2E1 activity in humans. METHODS: Healthy, male Caucasians ( n=19) were included. The multi-sample fractional clearance (Cl(fe)) of chlorzoxazone was compared with one...... estimates, Cl(est) at 3 h or 6 h, and MR at 3 h, can serve as reliable markers of CYP2E1 activity. The one-sample clearance method is an accurate, renal function-independent measure of the intrinsic activity; it is simple to use and easily applicable to humans.......-time-point clearance estimation (Cl(est)) at 3, 4, 5 and 6 h. Furthermore, the metabolite/drug ratios (MRs) estimated from one-time-point samples at 1, 2, 3, 4, 5 and 6 h were compared with Cl(fe). RESULTS: The concordance between Cl(est) and Cl(fe) was highest at 6 h. The minimal mean prediction error (MPE) of Cl...

  5. B(E1) Strengths from Coulomb Excitation of 11Be

    CERN Document Server

    Summers, N C; Ashwood, N I; Bouchat, V; Catford, W N; Clarke, N M; Curtis, N; Freer, M; Fulton, B R; Hanappe, F; Labiche, M; Lecouey, J L; Lemmon, R C; Mahboub, D; Ninane, A; Normand, G; Nunes, F M; Orr, N A; Pain, S D; Soic, N; Stuttgé, L; Thompson, I J; Timis, C N; Winfield, J S; Ziman, V

    2007-01-01

    The $B$(E1;$1/2^+\\to1/2^-$) strength for $^{11}$Be has been extracted from intermediate energy Coulomb excitation measurements, over a range of beam energies using a new reaction model, the extended continuum discretized coupled channels (XCDCC) method. In addition, a measurement of the excitation cross section for $^{11}$Be+$^{208}$Pb at 38.6 MeV/nucleon is reported. The $B$(E1) strength of 0.105(12) e$^2$fm$^2$ derived from this measurement is consistent with those made previously at 60 and 64 MeV/nucleon, i n contrast to an anomalously low result obtained at 43 MeV/nucleon. By coupling a multi-configuration description of the projectile structure with realistic reaction theory, the XCDCC model provides for the first time a fully quantum mechanical description of Coulomb excitation. The XCDCC calculations reveal that the excitation process involves significant contributions from nuclear, continuum, and higher-order effects. An analysis of the present and two earlier intermediate energy measurements yields a...

  6. SPAK and OSR1 Sensitive Cell Membrane Protein Abundance and Activity of KCNQ1/E1 K+ Channels

    Directory of Open Access Journals (Sweden)

    Bernat Elvira

    2015-11-01

    Full Text Available Background/Aims: KCNQ1/E1 channels are expressed in diverse tissues and serve a variety of functions including endolymph secretion in the inner ear, cardiac repolarization, epithelial transport and cell volume regulation. Kinases involved in regulation of epithelial transport and cell volume include SPAK (SPS1-related proline/alanine-rich kinase and OSR1 (oxidative stress-responsive kinase 1, which are under control of WNK (with-no-K[Lys] kinases. The present study explored whether KCNQ1/E1 channels are regulated by SPAK and/or OSR1. Methods: cRNA encoding KCNQ1/E1 was injected into Xenopus oocytes with or without additional injection of cRNA encoding wild-type SPAK, constitutively active T233ESPAK, WNK insensitive T233ASPAK, catalytically inactive D212ASPAK, wild-type OSR1, constitutively active T185EOSR1, WNK insensitive T185AOSR1 and catalytically inactive D164AOSR1. Voltage gated K+ channel activity was quantified utilizing dual electrode voltage clamp and KCNQ1/E1 channel protein abundance in the cell membrane utilizing chemiluminescence of KCNQ1/E1 containing an extracellular Flag tag epitope (KCNQ1-Flag/E1. Results: KCNQ1/E1 activity and KCNQ1-Flag/E1 protein abundance were significantly enhanced by wild-type SPAK and T233ESPAK, but not by T233ASPAK and D212ASPAK. Similarly, KCNQ1/E1 activity and KCNQ1-Flag/E1 protein abundance were significantly increased by wild-type OSR1 and T185EOSR1, but not by T185AOSR1 and D164AOSR1. Conclusions: SPAK and OSR1 participate in the regulation of KCNQ1/E1 protein abundance and activity.

  7. Cytotoxicity of acrylamide and its epoxide glycidamide in CHO cells expressing human cytochrome P450 2E1

    Institute of Scientific and Technical Information of China (English)

    Shoulin Wang; Xiaoyang He; Xinru Wang; Junyan Hong

    2006-01-01

    Objective: To investigate whether CYP2E1 is responsible for the acrylamide metabolic activation in Flp-In CHO cell system. Methods: CYP2E1 cDNA was subcloned from the human liver full-length cDNA library and subsequently transfected into the Flp-In CHO cells to generate the stable transfectant of CYP2E1. The CYP2E1 mRNA expression was determined by RT-PCR. Acrylamide and its epoxide glycidamide induced cytotoxicity and cell cycle arrest in G2/M were conducted using MTS assay and flow cytometry, respectively. Results: In the CHO cell stably expressing CYP2E1 (CHO-2E1), a ~1.5 kbsize of band was detected from the mRNA in the cells while no corresponding band in the CHO-vector cells, which indicated that CYP2E1 was successfully transfected in the CHO cells. Compared with the CHO-vector cells, acrylamide showed a concentrationdependent loss of viability in the CHO-2E1 cells but no significant change of G2/M arrest was found. As expected, glycidamide induced similar profile of cytotoxicity in both of the cells, and G2/M arrest presented a concentration-dependent increased in the CHO-2E1 cells. Conclusion: The result suggested that CYP2E1 might be responsible for the acrylamide metabolism, and its metabolite glycidamide was a direct cytotoxic and genotoxic agent. It should be further considered whether acrylamide-induced toxicity is through its epoxide glycidamide in the presence of CYP2E1.

  8. Crystal structure of hyperthermophilic esterase EstE1 and the relationship between its dimerization and thermostability properties

    Directory of Open Access Journals (Sweden)

    Koh Eunhee

    2007-07-01

    Full Text Available Abstract Background EstE1 is a hyperthermophilic esterase belonging to the hormone-sensitive lipase family and was originally isolated by functional screening of a metagenomic library constructed from a thermal environmental sample. Dimers and oligomers may have been evolutionally selected in thermophiles because intersubunit interactions can confer thermostability on the proteins. The molecular mechanisms of thermostabilization of this extremely thermostable esterase are not well understood due to the lack of structural information. Results Here we report for the first time the 2.1-Å resolution crystal structure of EstE1. The three-dimensional structure of EstE1 exhibits a classic α/β hydrolase fold with a central parallel-stranded beta sheet surrounded by alpha helices on both sides. The residues Ser154, Asp251, and His281 form the catalytic triad motif commonly found in other α/β hydrolases. EstE1 exists as a dimer that is formed by hydrophobic interactions and salt bridges. Circular dichroism spectroscopy and heat inactivation kinetic analysis of EstE1 mutants, which were generated by structure-based site-directed mutagenesis of amino acid residues participating in EstE1 dimerization, revealed that hydrophobic interactions through Val274 and Phe276 on the β8 strand of each monomer play a major role in the dimerization of EstE1. In contrast, the intermolecular salt bridges contribute less significantly to the dimerization and thermostability of EstE1. Conclusion Our results suggest that intermolecular hydrophobic interactions are essential for the hyperthermostability of EstE1. The molecular mechanism that allows EstE1 to endure high temperature will provide guideline for rational design of a thermostable esterase/lipase using the lipolytic enzymes showing structural similarity to EstE1.

  9. 26 CFR 1.501(e)-1 - Cooperative hospital service organizations.

    Science.gov (United States)

    2010-04-01

    ... anticipated needs of the organization. See, § 1.537-1(b). Whether there is an improper accumulation of funds... and operated by the United States, a State, the District of Columbia, or a possession of the...

  10. Hepatitis C Virus E1 and E2 Proteins Used as Separate Immunogens Induce Neutralizing Antibodies with Additive Properties.

    Science.gov (United States)

    Beaumont, Elodie; Roch, Emmanuelle; Chopin, Lucie; Roingeard, Philippe

    2016-01-01

    Various strategies involving the use of hepatitis C virus (HCV) E1 and E2 envelope glycoproteins as immunogens have been developed for prophylactic vaccination against HCV. However, the ideal mode of processing and presenting these immunogens for effective vaccination has yet to be determined. We used our recently described vaccine candidate based on full-length HCV E1 or E2 glycoproteins fused to the heterologous hepatitis B virus S envelope protein to compare the use of the E1 and E2 proteins as separate immunogens with their use as the E1E2 heterodimer, in terms of immunogenetic potential and the capacity to induce neutralizing antibodies. The specific anti-E1 and anti-E2 antibody responses induced in animals immunized with vaccine particles harboring the heterodimer were profoundly impaired with respect to those in animals immunized with particles harboring E1 and E2 separately. Moreover, the anti-E1 and anti-E2 antibodies had additive neutralizing properties that increase the cross-neutralization of heterologous strains of various HCV genotypes, highlighting the importance of including both E1 and E2 in the vaccine for an effective vaccination strategy. Our study has important implications for the optimization of HCV vaccination strategies based on HCV envelope proteins, regardless of the platform used to present these proteins to the immune system.

  11. Functional analysis of the C-terminal region of human adenovirus E1A reveals a misidentified nuclear localization signal

    Energy Technology Data Exchange (ETDEWEB)

    Cohen, Michael J.; King, Cason R.; Dikeakos, Jimmy D. [Department of Microbiology and Immunology, The University of Western Ontario, A4-833 London Regional Cancer Centre, 800 Commissioners Road E., London, Ontario, N6A 4L6 Canada (Canada); Mymryk, Joe S., E-mail: jmymryk@uwo.ca [Department of Microbiology and Immunology, The University of Western Ontario, A4-833 London Regional Cancer Centre, 800 Commissioners Road E., London, Ontario, N6A 4L6 Canada (Canada); Department of Oncology, The University of Western Ontario, London Regional Cancer Centre, Ontario (Canada)

    2014-11-15

    The immortalizing function of the human adenovirus 5 E1A oncoprotein requires efficient localization to the nucleus. In 1987, a consensus monopartite nuclear localization sequence (NLS) was identified at the C-terminus of E1A. Since that time, various experiments have suggested that other regions of E1A influence nuclear import. In addition, a novel bipartite NLS was recently predicted at the C-terminal region of E1A in silico. In this study, we used immunofluorescence microscopy and co-immunoprecipitation analysis with importin-α to verify that full nuclear localization of E1A requires the well characterized NLS spanning residues 285–289, as well as a second basic patch situated between residues 258 and 263 ({sup 258}RVGGRRQAVECIEDLLNEPGQPLDLSCKRPRP{sup 289}). Thus, the originally described NLS located at the C-terminus of E1A is actually a bipartite signal, which had been misidentified in the existing literature as a monopartite signal, altering our understanding of one of the oldest documented NLSs. - Highlights: • Human adenovirus E1A is localized to the nucleus. • The C-terminus of E1A contains a bipartite nuclear localization signal (NLS). • This signal was previously misidentified to be a monopartite NLS. • Key basic amino acid residues within this sequence are highly conserved.

  12. Hepatitis C Virus E1 and E2 Proteins Used as Separate Immunogens Induce Neutralizing Antibodies with Additive Properties

    Science.gov (United States)

    Beaumont, Elodie; Roch, Emmanuelle; Chopin, Lucie; Roingeard, Philippe

    2016-01-01

    Various strategies involving the use of hepatitis C virus (HCV) E1 and E2 envelope glycoproteins as immunogens have been developed for prophylactic vaccination against HCV. However, the ideal mode of processing and presenting these immunogens for effective vaccination has yet to be determined. We used our recently described vaccine candidate based on full-length HCV E1 or E2 glycoproteins fused to the heterologous hepatitis B virus S envelope protein to compare the use of the E1 and E2 proteins as separate immunogens with their use as the E1E2 heterodimer, in terms of immunogenetic potential and the capacity to induce neutralizing antibodies. The specific anti-E1 and anti-E2 antibody responses induced in animals immunized with vaccine particles harboring the heterodimer were profoundly impaired with respect to those in animals immunized with particles harboring E1 and E2 separately. Moreover, the anti-E1 and anti-E2 antibodies had additive neutralizing properties that increase the cross-neutralization of heterologous strains of various HCV genotypes, highlighting the importance of including both E1 and E2 in the vaccine for an effective vaccination strategy. Our study has important implications for the optimization of HCV vaccination strategies based on HCV envelope proteins, regardless of the platform used to present these proteins to the immune system. PMID:26966906

  13. 26 CFR 1.1033(e)-1 - Sale or exchange of livestock solely on account of drought.

    Science.gov (United States)

    2010-04-01

    ... of drought. 1.1033(e)-1 Section 1.1033(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... Sale or exchange of livestock solely on account of drought. (a) The sale or exchange of livestock... if the sale or exchange of such livestock by the taxpayer is solely on account of drought....

  14. Hepatitis C Virus E1 and E2 Proteins Used as Separate Immunogens Induce Neutralizing Antibodies with Additive Properties.

    Directory of Open Access Journals (Sweden)

    Elodie Beaumont

    Full Text Available Various strategies involving the use of hepatitis C virus (HCV E1 and E2 envelope glycoproteins as immunogens have been developed for prophylactic vaccination against HCV. However, the ideal mode of processing and presenting these immunogens for effective vaccination has yet to be determined. We used our recently described vaccine candidate based on full-length HCV E1 or E2 glycoproteins fused to the heterologous hepatitis B virus S envelope protein to compare the use of the E1 and E2 proteins as separate immunogens with their use as the E1E2 heterodimer, in terms of immunogenetic potential and the capacity to induce neutralizing antibodies. The specific anti-E1 and anti-E2 antibody responses induced in animals immunized with vaccine particles harboring the heterodimer were profoundly impaired with respect to those in animals immunized with particles harboring E1 and E2 separately. Moreover, the anti-E1 and anti-E2 antibodies had additive neutralizing properties that increase the cross-neutralization of heterologous strains of various HCV genotypes, highlighting the importance of including both E1 and E2 in the vaccine for an effective vaccination strategy. Our study has important implications for the optimization of HCV vaccination strategies based on HCV envelope proteins, regardless of the platform used to present these proteins to the immune system.

  15. CYP2E1 impairs GLUT4 gene expression and function: NRF2 as a possible mediator.

    Science.gov (United States)

    Armoni, M; Harel, C; Ramdas, M; Karnieli, E

    2014-06-01

    Impaired GLUT4 function/expression in insulin target tissues is well-documented in diabetes and obesity. Cytochrome P450 isoform 2E1 (CYP2E1) induces oxidative stress, leading to impaired insulin action. CYP2E1 knockout mice are protected against high fat diet-induced insulin resistance and obesity; however the molecular mechanisms are still unclear. We examined whether CYP2E1 impairs GLUT4 gene expression and function in adipose and muscle cells. CYP2E1 overexpression in skeletal muscle-derived L6 cells inhibited insulin-stimulated Glut4 translocation and 2-deoxyglucose uptake, with the latter inhibition being blocked by vitamin E. CYP2E1 overexpression in L6 and primary rat adipose (PRA) cells suppressed GLUT4 gene expression at promoter and mRNA levels, whereas CYP2E1 silencing had opposite effects. In PRA, CYP2E1-induced suppression of GLUT4 expression was blocked by chlormethiazole (CYP2E1-specific inhibitor) and the antioxidants vitamin E and N-acetyl-l-cysteine. CYP2E1 effect was mediated by the transcription factor NF-E2-related factor 2 (NRF2), as evident from its complete reversal by a coexpressed dominant-negative, but not wild-type NRF2. GLUT4 transcription was suppressed by NRF2 overexpression, and enhanced by NRF2 silencing. Promoter and ChIP analysis showed a direct and specific binding of NRF2 to a 58-326 GLUT4 promoter region that was required to maintain CYP2E1 suppression; this binding was enhanced by CYP2E1 overexpression. We suggest a mechanism for CYP2E1 action that involves: a) suppression of GLUT4 gene expression that is mediated by NRF2; b) impairment of insulin-stimulated Glut4 translocation and function. CYP2E1 and NRF2 are introduced as negative regulators of GLUT4 expression and function in insulin-sensitive cells.

  16. The hazard of the counterfeit E 1' signal in quartz to the ESR dating of fault movements

    Science.gov (United States)

    Toyoda, Shin; Schwarcz, Henry P.

    We have previously found (Toyoda and Schwarcz, in press) that γ-ray irradiation of granitic quartz produces a distinct ESR signal which overlaps the E 1' centre signal; this signal is removed by a heat treatment at 170°C for 15 min. Here we report the presence of this 'counterfeit' E 1[' signal in irradiated samples from fault gouge taken from the Hayward fault California. The contribution of the counterfeit E 1' signal to the total apparent E 1' centre signal intensity is significant and appears to account for most of the growth of the combined signal with irradiation. The intensity of the 'real' E 1' centre signal rises abruptly between irradiation doses of 300 and 1000 Gy, after removing the contribution of the counterfeit signal by heating.

  17. Evidence for the two-body nature of the E1 transition operator in the sdf-interacting boson model

    Energy Technology Data Exchange (ETDEWEB)

    Barfield, A.F.; Brentano, P. von; Dewald, A.; Zell, K.O.; Zamfir, N.V.; Bucurescu, D.; Ivascu, M.; Scholten, O.

    1989-01-01

    Two new a absolute transition rates are reported for the nucleus /sup 144/Sm following an (..cap alpha.., ..cap alpha..') Coulomb excitation study. They are B(E3;3/sup -/ -> 0/sup +/)=(38+-3)W.u. and B(E1;3/sup -/ -> 2/sup +/)=2.8+-0.4)x10/sup -3/W.u. This large E1 matrix element, along with the previously known B(E1;1/sup -/ -> 0/sup +/) value support the interpretation of the 1/sup -/ state in this nucleus as 2-phonon 2/sup +/x3/sup -/ excitation. In the frame of the IBM-1+f-boson model we show the need for a two-body term in the E1 transition operator. Estimates for the strenghts of the one and two-body parts of the E1 transition operator are obtained from these experimental data.

  18. Mammalian cytochrome CYP2E1 triggered differential gene regulation in response to trichloroethylene (TCE) in a transgenic poplar.

    Science.gov (United States)

    Kang, Jun Won; Wilkerson, Hui-Wen; Farin, Federico M; Bammler, Theo K; Beyer, Richard P; Strand, Stuart E; Doty, Sharon L

    2010-08-01

    Trichloroethylene (TCE) is an important environmental contaminant of soil, groundwater, and air. Studies of the metabolism of TCE by poplar trees suggest that cytochrome P450 enzymes are involved. Using poplar genome microarrays, we report a number of putative genes that are differentially expressed in response to TCE. In a previous study, transgenic hybrid poplar plants expressing mammalian cytochrome P450 2E1 (CYP2E1) had increased metabolism of TCE. In the vector control plants for this construct, 24 h following TCE exposure, 517 genes were upregulated and 650 genes were downregulated over 2-fold when compared with the non-exposed vector control plants. However, in the transgenic CYP2E1 plant, line 78, 1,601 genes were upregulated and 1,705 genes were downregulated over 2-fold when compared with the non-exposed transgenic CYP2E1 plant. It appeared that the CYP2E1 transgenic hybrid poplar plants overexpressing mammalian CYP2E1 showed a larger number of differentially expressed transcripts, suggesting a metabolic pathway for TCE to metabolites had been initiated by activity of CYP2E1 on TCE. These results suggest that either the over-expression of the CYP2E1 gene or the abundance of TCE metabolites from CYP450 2E1 activity triggered a strong genetic response to TCE. Particularly, cytochrome p450s, glutathione S-transferases, glucosyltransferases, and ABC transporters in the CYP2E1 transgenic hybrid poplar plants were highly expressed compared with in vector controls.

  19. MicroRNA-16 Modulates HuR Regulation of Cyclin E1 in Breast Cancer Cells

    Directory of Open Access Journals (Sweden)

    Xun Guo

    2015-03-01

    Full Text Available RNA binding protein (RBPs and microRNAs (miRNAs or miRs are post-transcriptional regulators of gene expression that are implicated in development of cancers. Although their individual roles have been studied, the crosstalk between RBPs and miRNAs is under intense investigation. Here, we show that in breast cancer cells, cyclin E1 upregulation by the RBP HuR is through specific binding to regions in the cyclin E1 mRNA 3' untranslated region (3'UTR containing U-rich elements. Similarly, miR-16 represses cyclin E1, dependent on its cognate binding sites in the cyclin E1 3'UTR. Evidence in the literature indicates that HuR can regulate miRNA expression and recruit or dissociate RNA-induced silencing complexes (RISC. Despite this, miR-16 and HuR do not affect the other’s expression level or binding to the cyclin E1 3'UTR. While HuR overexpression partially blocks miR-16 repression of a reporter mRNA containing the cyclin E1 3'UTR, it does not block miR-16 repression of endogenous cyclin E1 mRNA. In contrast, miR-16 blocks HuR-mediated upregulation of cyclin E1. Overall our results suggest that miR-16 can override HuR upregulation of cyclin E1 without affecting HuR expression or association with the cyclin E1 mRNA.

  20. Inhibition of human papillomavirus DNA replication by an E1-derived p80/UAF1-binding peptide.

    Science.gov (United States)

    Lehoux, Michaël; Fradet-Turcotte, Amélie; Lussier-Price, Mathieu; Omichinski, James G; Archambault, Jacques

    2012-04-01

    The papillomavirus E1 helicase is recruited by E2 to the viral origin, where it assembles into a double hexamer that orchestrates replication of the viral genome. We previously identified the cellular WD40 repeat-containing protein p80/UAF1 as a novel interaction partner of E1 from anogenital human papillomavirus (HPV) types. p80 was found to interact with the first 40 residues of HPV type 31 (HPV31) E1, and amino acid substitutions within this domain abrogated the maintenance of the viral episome in keratinocytes. In this study, we report that these p80-binding substitutions reduce by 70% the ability of E1 to support transient viral DNA replication without affecting its interaction with E2 and assembly at the origin in vivo. Microscopy studies revealed that p80 is relocalized from the cytoplasm to discrete subnuclear foci by E1 and E2. Chromatin immunoprecipitation assays further revealed that p80 is recruited to the viral origin in an E1- and E2-dependent manner. Interestingly, overexpression of a 40-amino-acid-long p80-binding peptide, derived from HPV31 E1, was found to inhibit viral DNA replication by preventing the recruitment of endogenous p80 to the origin. Mutant peptides defective for p80 interaction were not inhibitory, demonstrating the specificity of this effect. Characterization of this E1 peptide by nuclear magnetic resonance (NMR) showed that it is intrinsically disordered in solution, while mapping studies indicated that the WD repeats of p80 are required for E1 interaction. These results provide additional evidence for the requirement for p80 in anogenital HPV DNA replication and highlight the potential of E1-p80 interaction as a novel antiviral target.

  1. Energies and E1, M2 transition rates for Mo XXX

    CERN Document Server

    Hu, Feng; Mei, Maofei; Yang, Jiamin

    2016-01-01

    Based on relativistic wavefunctions from multiconfigurational Dirac-Hartree-Fock (MCDHF) and configuration interaction calculations, energy levels, radiative rates, and wavelengths are evaluated for all levels of 3s$^2$3p, 3s3p$^2$, 3s$^2$3d, 3p$^3$, 3s3p3d, 3p$^2$3d and 3s3d$^2$ configurations of Al-like Molybdenum ion (Mo XXX). Transition probabilities are reported for E1 and M2 transitions from the ground level. The valence-valence and core-valence correlation effects are accounted for in a systematic way. Breit interactions and quantum electrodynamics effects are estimated in subsequent relativistic configuration interaction calculations. Comparisons are made with the available data in the literature and good agreement has been found which confirms the reliability of our results.

  2. Chang'E-1 orbiter discovers a lunar nearside volcano: YUTU Mountain

    Institute of Scientific and Technical Information of China (English)

    PING dinSong; HUANG Qian; SU XiaoLi; TANG GeShi; SHU Rong; XIAO Long; HUANG Jun

    2009-01-01

    In the day time of the Moon surface, the strong illumination from high altitude and high albedo rate radical craters will introduce the illumination effect on observing the nearby low altitude, low albedo rate and shallow small slop rate area seriously, and even can "hide" the later area from the light. Based on the lunar global topography model obtained by Chang'E-1 mission, and by comparing with the lunar gravity model, a volcano named "YUTU Mountain" has been identified. It is a volcano with diameter of ~300 km and height of ~2 km located at (14°N, 308°E) in Oceanus Procellarum. Besides, the DEM of another volcano named "GUISHU Mountain" in the same area has been improved. This new discovery will benefit the study of lunar magmatism and volcanism evolution in the nearside of the Moon.

  3. The use of laser altimetry data in Chang'E-1 precision orbit determination

    Science.gov (United States)

    Chang, Sheng-Qi; Huang, Yong; Li, Pei-Jia; Hu, Xiao-Gong; Fan, Min

    2016-09-01

    Accurate altimetric measurement not only can be applied to the calculation of a topography model but also can be used to improve the quality of the orbit reconstruction in the form of crossovers. Altimetry data from the Chang'E-1 (CE-1) laser altimeter are analyzed in this paper. The differences between the crossover constraint equation in the form of height discrepancies and in the form of minimum distances are mainly discussed. The results demonstrate that the crossover constraint equation in the form of minimum distances improves the CE-1 orbit precision. The overlap orbit performance has increased ∼ 30% compared to the orbit using only tracking data. External assessment using the topography model also shows orbit improvement. The results will be helpful for recomputing ephemeris and improving the CE-1 topography model.

  4. Application of ozonation process in industrial wastewaters: textile, kraft E1 and whey effluents.

    Science.gov (United States)

    Assalin, M R; Almeida, E S; Rosa, M A; Moraes, S G; Duran, N

    2004-08-01

    A large variety of organic and inorganic compounds can be found in wastewater from industrial processes. In this work, Advanced Oxidative Processes (AOPs) have been applied for the control of water pollution and the ozonation of different effluents was investigated. Wastewater from textile, kraft E1 and cheese manufacturing processes were chosen as examples of industrial effluents. The efficiency of substrate mineralization has been comparatively analyzed by the decrease in total organic carbon (TOC), color, and toxicity. The results revealed that the ozonation process can be a method for decolorization of effluent, but it is not effective for TOC reduction. The whey effluent was the most recalcitrant wastewater for ozone treatment which produced no TOC removal.

  5. Crystal structure of (E-1-(4-chlorophenylethanone O-dehydroabietyloxime

    Directory of Open Access Journals (Sweden)

    Jian-Qiang Zheng

    2014-09-01

    Full Text Available The title compound, C28H34ClNO2 {systematic name: (E-1-(4-chlorophenylethanone O-[(1R,4aS,10aR-7-isopropyl-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]oxime}, was synthesized from dehydroabietic acid. In the dehydroabietyl moiety, the central and terminal cyclohexane rings display chair and half-chair conformations, respectively, and a trans-ring junction. The C=N bond is in an E conformation and the C—O—N=C torsion angle is 148.1 (5°. No directional interactions except van der Waals contacts occur in the crystal structure.

  6. Lunar digital elevation model and elevation distribution model based on Chang’E-1 LAM data

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    More than 8.2 million effective data samples were obtained by the Chang’E-1 Laser Altimeter (LAM).In order to produce a global topographic model of the moon with improved accuracy,a hierarchical many-knot spline method was proposed in this paper.This algorithm makes use of a hierarchy of control lattices to approximate or interpolate the LAM data.Based on the proposed algorithm,a 0.0625°×0.0625° grid of global lunar DEM was obtained and it was compared with ULCN2005,CLTMs01 and Kaguya models,respectively.At the same time,this paper explored the elevation distribution law and established the elevation distribution model.It is shown that the global lunar and nearside elevation distribution is positively skewed and leptokurtic normal distribution,and the farside elevation distribution is a positively skewed and platykurtic normal distribution.

  7. The use of laser altimetry data in Chang'E-1 precision orbit determination

    Science.gov (United States)

    Chang, Sheng-Qi; Huang, Yong; Li, Pei-Jia; Hu, Xiao-Gong; Fan, Min

    2016-09-01

    Accurate altimetric measurement not only can be applied to the calculation of a topography model but also can be used to improve the quality of the orbit reconstruction in the form of crossovers. Altimetry data from the Chang'E-1 (CE-1) laser altimeter are analyzed in this paper. The differences between the crossover constraint equation in the form of height discrepancies and in the form of minimum distances are mainly discussed. The results demonstrate that the crossover constraint equation in the form of minimum distances improves the CE-1 orbit precision. The overlap orbit performance has increased ˜ 30% compared to the orbit using only tracking data. External assessment using the topography model also shows orbit improvement. The results will be helpful for recomputing ephemeris and improving the CE-1 topography model.

  8. Melatonin Suppresses Autophagy Induced by Clinostat in Preosteoblast MC3T3-E1 Cells.

    Science.gov (United States)

    Yoo, Yeong-Min; Han, Tae-Young; Kim, Han Sung

    2016-04-08

    Microgravity exposure can cause cardiovascular and immune disorders, muscle atrophy, osteoporosis, and loss of blood and plasma volume. A clinostat device is an effective ground-based tool for simulating microgravity. This study investigated how melatonin suppresses autophagy caused by simulated microgravity in preosteoblast MC3T3-E1 cells. In preosteoblast MC3T3-E1 cells, clinostat rotation induced a significant time-dependent increase in the levels of the autophagosomal marker microtubule-associated protein light chain (LC3), suggesting that autophagy is induced by clinostat rotation in these cells. Melatonin treatment (100, 200 nM) significantly attenuated the clinostat-induced increases in LC3 II protein, and immunofluorescence staining revealed decreased levels of both LC3 and lysosomal-associated membrane protein 2 (Lamp2), indicating a decrease in autophagosomes. The levels of phosphorylation of mammalian target of rapamycin (p-mTOR) (Ser2448), phosphorylation of extracellular signal-regulated kinase (p-ERK), and phosphorylation of serine-threonine protein kinase (p-Akt) (Ser473) were significantly reduced by clinostat rotation. However, their expression levels were significantly recovered by melatonin treatment. Also, expression of the Bcl-2, truncated Bid, Cu/Zn- superoxide dismutase (SOD), and Mn-SOD proteins were significantly increased by melatonin treatment, whereas levels of Bax and catalase were decreased. The endoplasmic reticulum (ER) stress marker GRP78/BiP, IRE1α, and p-PERK proteins were significantly reduced by melatonin treatment. Treatment with the competitive melatonin receptor antagonist luzindole blocked melatonin-induced decreases in LC3 II levels. These results demonstrate that melatonin suppresses clinostat-induced autophagy through increasing the phosphorylation of the ERK/Akt/mTOR proteins. Consequently, melatonin appears to be a potential therapeutic agent for regulating microgravity-related bone loss or osteoporosis.

  9. Study of thermally excited nuclei through E1 and E2 decay from collective modes

    Energy Technology Data Exchange (ETDEWEB)

    Million, B.; Bracco, A.; Benzoni, G.; Leoni, S.; Camera, F.; Wieland, O. [Dipartimento di Fisica, Universita di Milano and INFN, via Celoria 16, 20133, Milano (Italy); Maj, A.; Kmiecik, M. [Niewodniczanski Institute of Nuclear Physics, 31-342, Krakow (Poland); Gadea, A. [Laboratori Nazionali di Legnaro, via Romea, Legnaro (Italy); Herskind, B. [The Niels Bohr Institute, Blegdamsvej 15-17, 2100, Copenhagen (Denmark)

    2004-04-01

    The nuclear system at the limit of excitation energy and angular momentum is here studied in the case of the superdeformed nucleus {sup 143}Eu using {gamma}-spectroscopy techniques. The data are based on a EUROBALL experiment using the reaction {sup 37}Cl+{sup 110}Pd{yields}{sup 143}Eu+4n. The influence of thermal energy on superdeformed configurations is investigated through the analysis of the quasi-continuum spectra formed by E2 transitions among states of excited rotational bands with energy extending up to 4-5 MeV above the yrast line. In particular, the effective lifetimes of the discrete rotational bands forming ridge structures in {gamma}-{gamma} coincidence matrices is measured by a Doppler Shift Attenuation Method. The deduced quadrupole deformation of Q{sub t} {approx}10 eb indicates that the nucleus maintains its collectivity with increasing excitation energy, supporting the superdeformed character of the excited nuclear rotation. The obtained number of superdeformed discrete bands forming the ridge structures is found in good agreement with microscopic cranked shell model calculations including the decay-out process into the lower deformation minimum. In addition, the nuclear properties at higher excitation energies are investigated through the E1 {gamma}-decay of the giant dipole resonance (GDR). It is found that the intensity of the superdeformed yrast and excited bands increases by a factor of approximately 1.6 when a coincidence with a high-energy {gamma}-ray is required, showing the importance of the E1 cooling in the feeding mechanism of the superdeformed states. (orig.)

  10. (E-1-(1,3-Benzodioxol-5-yl-3-(3-bromophenylprop-2-en-1-one

    Directory of Open Access Journals (Sweden)

    H. S. Yathirajan

    2008-12-01

    Full Text Available In the title compound, C16H11BrO3, the molecules adopt an E configuration with respect to the C=C double bond of the propenone unit. The 13 non-H atoms of the benzodioxole and propenone units are approximately coplanar (r.m.s. deviation = 0.027 Å and the bromobenzene ring plane forms a dihedral angle of 10.8 (1° to this plane. The structure is layered, with the molecules forming a herring-bone arrangement within each layer.

  11. (E-1-(4-Chlorophenyl-3-[4-(dimethylaminophenyl]prop-2-en-1-one

    Directory of Open Access Journals (Sweden)

    Xinyou Lei

    2009-04-01

    Full Text Available The title compound, C17H16ClNO, was synthesized using a solvent-free method by reaction of 4-(dimethylaminobenzaldehyde with 4-chloroacetophenone and NaOH. The chlorophenyl ring makes a dihedral angle of 18.1 (3° with the central propenone unit, while the (dimethylaminophenyl group is disordered over two orientations of equal occupancies, which make dihedral angles with the central propenone unit of 32.9 (3 and 57.4 (3°, respectively.

  12. Chromosomal damage induced by human adenovirus type 12 requires expression of the E1B 55-kilodalton viral protein.

    Science.gov (United States)

    Schramayr, S; Caporossi, D; Mak, I; Jelinek, T; Bacchetti, S

    1990-01-01

    Infection of human embryonic kidney cells with adenovirus type 12 results in the induction of damage at specific (17q21-22, 1p36, 1q21, and 1q42-43) and random sites in the cellular chromosomes. A previous study by Durnam et al. (D. M. Durnam, P. P. Smith, J. C. Menninger, and J. K. McDougall, Cancer Cells 4:349-354, 1986) indicated that the expression of viral early region 1 (E1) is sufficient for the induction of damage at band 17q21-22. In the present report we used an adenovirus type 12-adenovirus type 5 recombinant with E1A hybrid sequences as well as viruses with mutations in the adenovirus type 12 E1B genes to map adenovirus type 12 E1 functions involved in the induction of genetic damage. Our results show that the expression of the E1A proteins is not sufficient for this effect. On the other hand, mutations within the E1B 55-kilodalton protein but not the E1B 19-kilodalton protein affect the ability of the virus to induce both specific and random chromosomal damage. Images PMID:2325204

  13. Adenovirus type 12 E1B 55-kilodalton oncoprotein promotes p53-mediated apoptotic response of ovarian cancer to cisplatin.

    Science.gov (United States)

    Wang, Junnai; Gao, Qinglei; Li, Qiang

    2015-08-01

    The tumor suppressor p53-mediated apoptotic response plays an important role in cisplatin resistant in ovarian cancer. The adenovirus (Ad) type 12 E1B 55-kDa protein binds to p53 and inactivates its transcriptional transactivation function. In this study, we test the hypothesis that Ad12 E1B 55-kDa oncoprotein promotes p53-mediated apoptotic response of ovarian cancer to cisplatin. First, we observed the upregulation protein level of p53 target genes in cisplatin-resistant or cisplatin-sensitive ovarian cancer by Western blotting. Second, after transfection of Ad12 E1b 55-kDa expression plasmid, the expressions of p53 target genes in A2780 cells were further enhanced. Co-IP experiment demonstrated Ad12 E1b 55 kDa associated with p53. MTT assay confirmed that the cell proliferation was enhanced after transfection, as well as the enhanced cell inhibitory rate in the presence of cisplatin. Using flow cytometry, transfection of Ad12 E1B 55-kDa protein induced apoptosis and promoted S-phase transition in proliferation. Finally, results showed that all these changes promoted by Ad12 E1b 55 kDa were attenuated by the exposure of specific inhibitor of p53 signaling, pifithrin-α. Taken together, we concluded that Ad E1B 55-kDa oncoprotein promotes p53-mediated apoptotic response of ovarian cancer to cisplatin.

  14. Intranuclear targeting and nuclear export of the adenovirus E1B-55K protein are regulated by SUMO1 conjugation

    OpenAIRE

    Kindsmüller, Kathrin; Groitl, Peter; Härtl, Barbara; Blanchette, Paola; Hauber, Joachim; Dobner, Thomas

    2007-01-01

    We have investigated the requirements for CRM1-mediated nuclear export and SUMO1 conjugation of the adenovirus E1B-55K protein during productive infection. Our data show that CRM1 is the major export receptor for E1B-55K in infected cells. Functional inactivation of the E1B-55K CRM1-dependent nuclear export signal (NES) or leptomycin B treatment causes an almost complete redistribution of the viral protein from the cytoplasm to the nucleus and its accumulation at the periphery of the viral re...

  15. H20-20 E1远端模块在调度系统中的应用

    Institute of Scientific and Technical Information of China (English)

    欧阳雪松; 陈立新

    2005-01-01

    文章主要针对H20-20交换机系统下的E1远端模块进行了详细的描述,阐述E1远端模块的性能特点和联接方式.重点介绍了E1远端模块在电力调度系统中的实际应用,以及在和PDH/SDH设备联接时时钟同步问题的解决方案.

  16. CYP2E1 Pst Ⅰ/Rsa Ⅰ polymorphism and colorectal cancer risk:A meta-analysis

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    AIM:To clarify the association between CYP2E1 PstⅠ/RsaⅠ polymorphism and susceptibility to colorectal cancer.METHODS:A meta-analysis based on 10 eligible casecontrol studies involving 4979 cases and 6012 controls was carried out to summarize the data on the association between CYP2E1 RsaⅠ/PstⅠ polymorphism and colorectal cancer risk.RESULTS:In comparison of the homozygote c2c2 and c2 carriers(c1c2 + c2c2) and the homozygous wild-type genotype(c1c1),no association was found between CYP2E1 RsaⅠ/PstⅠ polymorph...

  17. High-yield expression in Escherichia coli and purification of mouse ubiquitin-activating enzyme E1.

    Science.gov (United States)

    Carvalho, Andreia F; Pinto, Manuel P; Grou, Cláudia P; Vitorino, Rui; Domingues, Pedro; Yamao, Fumiaki; Sá-Miranda, Clara; Azevedo, Jorge E

    2012-07-01

    Research in the ubiquitin field requires large amounts of ubiquitin-activating enzyme (E1) for in vitro ubiquitination assays. Typically, the mammalian enzyme is either isolated from natural sources or produced recombinantly using baculovirus/insect cell protein expression systems. Escherichia coli is seldom used to produce mammalian E1 probably due to the instability and insolubility of this high-molecular mass protein. In this report, we show that 5-10 mg of histidine-tagged mouse E1 can be easily obtained from a 1 l E. coli culture. A low temperature during the protein induction step was found to be critical to obtain an active enzyme.

  18. Occupational Toluene Exposure Induces Cytochrome P450 2E1 mRNA Expression in Peripheral Lymphocytes

    OpenAIRE

    Mendoza-Cantú, Ania; Castorena-Torres, Fabiola; León,Mario Bermúdez de; Cisneros, Bulmaro; López-Carrillo, Lizbeth; Rojas-García, Aurora E.; Aguilar-Salinas, Alberto; Manno, Maurizio; Albores, Arnulfo

    2005-01-01

    Print workers are exposed to organic solvents, of which the systemic toxicant toluene is a main component. Toluene induces expression of cytochrome P450 2E1 (CYP2E1), an enzyme involved in its own metabolism and that of other protoxicants, including some procarcinogens. Therefore, we investigated the association between toluene exposure and the CYP2E1 response, as assessed by mRNA content in peripheral lymphocytes or the 6-hydroxychlorzoxazone (6OH-CHZ)/chlorzoxazone (CHZ) quotient (known as ...

  19. 两类E1 Nino事件与杭州气候的相关性分析

    Institute of Scientific and Technical Information of China (English)

    周蕾芝

    2002-01-01

    根据赤道东太平洋海温开始增温位置不同,可将E1 Nino事件分为两类.本文运用x2检验分析这两类E1 Nino事件与杭州降水、气温的相关关系.分析结果表明,两类E1 Nin0事件与杭州汛期降水和全年降水量以及7月平均气温关系密切.

  20. Human ubiquitin-activating enzyme, E1. Indication of potential nuclear and cytoplasmic subpopulations using epitope-tagged cDNA constructs.

    Science.gov (United States)

    Handley-Gearhart, P M; Stephen, A G; Trausch-Azar, J S; Ciechanover, A; Schwartz, A L

    1994-12-30

    The ubiquitin-activating enzyme E1 catalyzes the first step in the ubiquitin conjugation pathway. Previously, we have cloned and sequenced the cDNA for human E1. Expression of the E1 cDNA in the ts20 cell line, which harbors a thermolabile E1, abrogated the phenotypic defects associated with this line. However, little is known of the cell biology of the E1 protein or the nature of the E1 doublet. Thus, we constructed epitope-tagged E1 cDNAs in which the HA monoclonal antibody epitope tag sequence (from influenza hemagglutinin and recognized by the 12CA5 monoclonal antibody) was fused to the amino terminus of E1. Because the amino-terminal amino acid sequence of E1 is unknown, three constructs were made in which the HA tag was placed at each of the first three ATGs in the open reading frame (HA-1E1, HA-2E1, and HA-3E1). Western analysis of HeLa cells transfected with the constructs revealed that HA-1E1 closely comigrated with the upper band of the E1 doublet, and HA-2E1 comigrated with the lower band of the E1 doublet; HA-3E1 appeared smaller than either of the E1 bands. Metabolic labeling with 32P and immunoprecipitation with anti-HA antibody revealed that only the HA-1E1 protein product is phosphorylated; polyclonal anti-E1 antibody showed that only the upper band of the endogenous E1 doublet is phosphorylated. Each of the constructs was able to rescue the mutant phenotype of the ts20 cell line. Immunofluorescence studies showed that HA-2E1 and HA-3E1 were distributed in the cytoplasm with both negative and positive nuclei. This pattern of distribution has also been observed when immunostaining with a monoclonal antibody to E1 (1C5). However, the staining pattern associated with a polyclonal anti-E1 antibody (JJJ) is characterized by positive staining cytoplasm and nuclei in all cells. The HA-1E1 construct exhibited apparently exclusive nuclear distribution in HeLa cells. The difference between the staining patterns of the polyclonal and monoclonal anti-E1

  1. ColE1-Plasmid Production in Escherichia coli: Mathematical Simulation and Experimental Validation

    Science.gov (United States)

    Freudenau, Inga; Lutter, Petra; Baier, Ruth; Schleef, Martin; Bednarz, Hanna; Lara, Alvaro R.; Niehaus, Karsten

    2015-01-01

    Plasmids have become very important as pharmaceutical gene vectors in the fields of gene therapy and genetic vaccination in the past years. In this study, we present a dynamic model to simulate the ColE1-like plasmid replication control, once for a DH5α-strain carrying a low copy plasmid (DH5α-pSUP 201-3) and once for a DH5α-strain carrying a high copy plasmid (DH5α-pCMV-lacZ) by using ordinary differential equations and the MATLAB software. The model includes the plasmid replication control by two regulatory RNA molecules (RNAI and RNAII) as well as the replication control by uncharged tRNA molecules. To validate the model, experimental data like RNAI- and RNAII concentration, plasmid copy number (PCN), and growth rate for three different time points in the exponential phase were determined. Depending on the sampled time point, the measured RNAI- and RNAII concentrations for DH5α-pSUP 201-3 reside between 6 ± 0.7 and 34 ± 7 RNAI molecules per cell and 0.44 ± 0.1 and 3 ± 0.9 RNAII molecules per cell. The determined PCNs averaged between 46 ± 26 and 48 ± 30 plasmids per cell. The experimentally determined data for DH5α-pCMV-lacZ reside between 345 ± 203 and 1086 ± 298 RNAI molecules per cell and 22 ± 2 and 75 ± 10 RNAII molecules per cell with an averaged PCN of 1514 ± 1301 and 5806 ± 4828 depending on the measured time point. As the model was shown to be consistent with the experimentally determined data, measured at three different time points within the growth of the same strain, we performed predictive simulations concerning the effect of uncharged tRNA molecules on the ColE1-like plasmid replication control. The hypothesis is that these tRNA molecules would have an enhancing effect on the plasmid production. The in silico analysis predicts that uncharged tRNA molecules would indeed increase the plasmid DNA production. PMID:26389114

  2. ColE1-plasmid production in Escherichia coli: Mathematical Simulation and Experimental Validation

    Directory of Open Access Journals (Sweden)

    Inga eFreudenau

    2015-09-01

    Full Text Available Plasmids have become very important as pharmaceutical gene vectors in the fields of gene therapy and genetic vaccination in the last years. In this study, we present a dynamic model to simulate the ColE1-like plasmid replication control, once for a DH5α-strain carrying a low copy plasmid (DH5α-pSUP 201-3 and once for a DH5α-strain carrying a high copy plasmid (DH5α-pCMV-lacZ by using ordinary differential equations (ODE and the MATLAB software. The model includes the plasmid replication control by two regulatory RNA molecules (RNAI and RNAII as well as the replication control by uncharged tRNA molecules. To validate the model, experimental data like RNAI- and RNAII concentration, plasmid copy number (PCN, and growth rate for three different time points in the exponential phase were determined. Depending on the sampled time point, the measured RNAI and RNAII concentrations for DH5α-pSUP 201-3 reside between 6 ±0.7 to 34 ±7 RNAI molecules per cell and 0.44 ±0.1 to 3 ±0.9 RNAII molecules per cell. The determined plasmid copy numbers (PCN averaged between 46 ±26 to 48 ±30 plasmids per cell. The experimentally determined data for DH5α-pCMV-lacZ reside between 345 ±203 to 1086 ±298 RNAI molecules per cell and 22 ±2 to 75 ±10 RNAII molecules per cell with an averaged PCN of 1514 ±1301 to 5806 ±4828 depending on the measured time point. As the model was shown to be consistent with the experimentally determined data, measured at three different time points within the growth of the same strain, we performed predictive simulations concerning the effect of uncharged tRNA molecules on the ColE1-like plasmid replication control. The hypothesis is that these tRNA molecules would have an enhancing effect on the plasmid production. The in silico analysis predicts that uncharged tRNA molecules would indeed increase the pDNA production.

  3. Prokaryotic expression of E1 genes of bovine viral diarrhea virus and detection of the expressed protein%牦牛病毒性腹泻病毒E1基因的原核表达

    Institute of Scientific and Technical Information of China (English)

    王盼盼; 刘亚刚; 孙凯; 王文伯; 胡炳峰; 于吉锋; 冯英阳

    2010-01-01

    以牦牛BVDV基因组DNA为模板, 运用聚合酶链反应成功扩增出牦牛BVDV E1基因, 将其插入到pET-28a及pET-32a原核表达载体, 构建重组表达质粒pET-28a-E1和pET-32a-E1并分别转化至Rosetta(DE3)和BL21(DE3)宿主菌中, IPTG诱导表达, 经SDS-PAGE检测, pET-28a-E1和pET-32a-E1在宿主菌Rosetta(DE3)和BL21(DE3)中均表达出约40KU的融合蛋白, 结果分析表明E1基因在两个宿主中都获得了高效表达, 表达出的蛋白大小与预期结果相符.

  4. Generating Units

    Data.gov (United States)

    Department of Homeland Security — Generating Units are any combination of physically connected generators, reactors, boilers, combustion turbines, and other prime movers operated together to produce...

  5. Pharmacodynamic comparison of prostaglandin E1 administered by different routes to rats.

    Science.gov (United States)

    Gu, Fu-gen; Cui, Fu-de; Gao, Yong-liang

    2007-07-01

    The pharmacodynamics of prostaglandin E1 (PGE1) administered by different routes to rats was investigated in this paper. The hypotensive effect of PGE, was used as an index of drug efficacy, pharmacodynamic parameters such as time to reach peak effect (Tmax), maximal percentage of blood pressure decrease (Emax, %), duration of effect (Td), and the area under the blood pressure decrease percent-time curves (AUC, % x min) were determined after PGE1 given to rats intranasally, sublingually, intraperitoneally (ip), and intramuscularly (im), separately, and compared with those obtained from intravenous (iv) administration. Similar to iv route, the pharmacodynamic parameters of PGE1 from the other administration routes, Emax, Td and in particular AUC values were all increased with increasing doses, showing dose-efficacy relationship. Tmax was found to be approximately 3-4 min for nasal route, 3-8 min for im, 6-8 min for ip and 12-30 min for sublingual route, separately. Thus, the order of magnitude of absorption rate of the drug was as follows: nasal approximately = im > ip > sublingual. If the pharmacological bioavailability (PF) for each administration route was used as a tentative measure of drug absorption extent, the order of magnitude of absolute bioavailability appeared as follows: nasal > im approximately = ip > sublingual. Furthermore, the interindividual difference was found to be larger for im and ip route than that for nasal and sublingual route. These results indicate nasal and sublingual routes are two promising routes for the systemic delivery of PGE1 in clinical applications.

  6. Evaluation of Rational Function Model for Geometric Modeling of CHANG'E-1 CCD Images

    Science.gov (United States)

    Liu, Y.; Di, K.

    2011-08-01

    Rational Function Model (RFM) is a generic geometric model that has been widely used in geometric processing of high-resolution earth-observation satellite images, due to its generality and excellent capability of fitting complex rigorous sensor models. In this paper, the feasibility and precision of RFM for geometric modeling of China's Chang'E-1 (CE-1) lunar orbiter images is presented. The RFM parameters of forward-, nadir- and backward-looking CE-1 images are generated though least squares solution using virtual control points derived from the rigorous sensor model. The precision of the RFM is evaluated by comparing with the rigorous sensor model in both image space and object space. Experimental results using nine images from three orbits show that RFM can precisely fit the rigorous sensor model of CE-1 CCD images with a RMS residual error of 1/100 pixel level in image space and less than 5 meters in object space. This indicates that it is feasible to use RFM to describe the imaging geometry of CE-1 CCD images and spacecraft position and orientation. RFM will enable planetary data centers to have an option to supply RFM parameters of orbital images while keeping the original orbit trajectory data confidential.

  7. Dem Extraction from CHANG'E-1 Lam Data by Surface Skinning Technology

    Science.gov (United States)

    Zhang, X.-B.; Zhang, W.-M.

    2011-08-01

    DEM is a digital model or 3-D representation of a terrain's surface and it is created from terrain elevation data. The main models for DEM extraction based on Lidar data or Laser Altimeter data currently use the idea that point cloud is scattered, such as regular grid model, TIN model and contour model. Essentially, in these above methods, the discrete points are interpolated into regular grid data and irregular grid data. In fact, point cloud generated by Laser Altimeter is not totally scattered, but have some regularity. In this paper, to utilize this regularity, the proposed method adopts surface skinning technology to generate DEM from Chang'E-1 Laser Altimeter data. The surface skinning technology is widely used in the field of mechanical engineering. Surface skinning is the process of passing a smooth surface through a set of curves called sectional curves, which, in general, may not be compatible. In the process of generating section line, a need for attention is that it needs to use curvature method to get a set of characteristic points, and these feature points were used to subdivide segment; the next step is generating several curves on some key places. These curves describe the shape of the curved surface. The last step is to generate a curved surface that through these curves. The result shows that, this idea is feasible, useful and it provides a novel way to generate accurate DEM.

  8. Hierarchical polymeric scaffolds support the growth of MC3T3-E1 cells.

    Science.gov (United States)

    Akbarzadeh, Rosa; Minton, Joshua A; Janney, Cara S; Smith, Tyler A; James, Paul F; Yousefi, Azizeh-Mitra

    2015-02-01

    Tissue engineering makes use of the principles of biology and engineering to sustain 3D cell growth and promote tissue repair and/or regeneration. In this study, macro/microporous scaffold architectures have been developed using a hybrid solid freeform fabrication/thermally induced phase separation (TIPS) technique. Poly(lactic-co-glycolic acid) (PLGA) dissolved in 1,4-dioxane was used to generate a microporous matrix by the TIPS method. The 3D-bioplotting technique was used to fabricate 3D macroporous constructs made of polyethylene glycol (PEG). Embedding the PEG constructs inside the PLGA solution prior to the TIPS process and subsequent extraction of PEG following solvent removal (1,4-dioaxane) resulted in a macro/microporous structure. These hierarchical scaffolds with a bimodal pore size distribution (300 μm) contained orthogonally interconnected macro-channels generated by the extracted PEG. The diameter of the macro-channels was varied by tuning the dispensing parameters of the 3D bioplotter. The in vitro cell culture using murine MC3T3-E1 cell line for 21 days demonstrated that these scaffolds could provide a favorable environment to support cell adhesion and growth.

  9. Modulation of Osteogenesis in MC3T3-E1 Cells by Different Frequency Electrical Stimulation.

    Directory of Open Access Journals (Sweden)

    Yu Wang

    Full Text Available Electrical stimulation (ES is therapeutic to many bone diseases, from promoting fracture regeneration to orthopedic intervention. The application of ES offers substantial therapeutic potential, while optimal ES parameters and the underlying mechanisms responsible for the positive clinical impact are poorly understood. In this study, we assembled an ES cell culture and monitoring device. Mc-3T3-E1 cells were subjected to different frequency to investigate the effect of osteogenesis. Cell proliferation, DNA synthesis, the mRNA levels of osteosis-related genes, the activity of alkaline phosphatase (ALP, and intracellular concentration of Ca2+ were thoroughly evaluated. We found that 100 Hz could up-regulate the mRNA levels of collagen I, collagen II and Runx2. On the contrary, ES could down-regulate the mRNA levels of osteopontin (OPN. ALP activity assay and Fast Blue RR salt stain showed that 100 Hz could accelerate cells differentiation. Compared to the control group, 100 Hz could promote cell proliferation. Furthermore, 1 Hz to 10 Hz could improve calcium deposition in the intracellular matrix. Overall, these results indicate that 100Hz ES exhibits superior potentialities in osteogenesis, which should be beneficial for the clinical applications of ES for the treatment of bone diseases.

  10. MC3T3-E1 Cells on Titanium Surfaces with Nanometer Smoothness and Fibronectin Immobilization

    Directory of Open Access Journals (Sweden)

    Tohru Hayakawa

    2012-01-01

    Full Text Available The present study was aimed to evaluate the viability and total protein contents of osteoblast-like cells on the titanium surface with different surface mechanical treatment, namely, nanometer smoothing (Ra: approximately 2.0 nm and sandblasting (Ra: approximately 1.0 μm, and biochemical treatment, namely, with or without fibronectin immobilization. Fibronectin could be easily immobilized by tresyl chloride-activation technique. MC3T3-E1 cells were seeded on the different titanium surfaces. Cell viability was determined by MTT assay. At 1 day of cell culture, there were no significant differences in cell viability among four different titanium surfaces. At 11 days, sandblasted titanium surface with fibronectin immobilization showed the significantly highest cell viability than other titanium surface. No significant differences existed for total protein contents among four different titanium surfaces at 11 days of cell culture. Scanning electron microscopy observation revealed that smoothness of titanium surface produced more spread cell morphologies, but that fibronectin immobilization did not cause any changes of the morphologies of attached cells. Fibronectin immobilization provided greater amount of the number of attached cells and better arrangement of attached cells. In conclusion, the combination of sandblasting and fibronectin immobilization enhanced the cell viability and fibronectin immobilization providing better arrangements of attached cells.

  11. CYP2E1, oxidative stress, post-translational modifications and lipid metabolism.

    Science.gov (United States)

    Lakshman, M Raj; Garige, Mamatha; Gong, Maokai A; Leckey, Leslie; Varatharajalu, Ravi; Redman, Robert S; Seth, Devanshi; Haber, Paul S; Hirsch, Kenneth; Amdur, Richard; Shah, Ruchi

    2013-01-01

    Chronic alcohol-mediated down-regulation of hepatic ST6Gal1 gene leads to defective glycosylation of lipid-carrying apolipoproteins such as apo E and apo J, resulting in defective VLDL assembly and intracellular lipid and lipoprotein transport, which in turn is responsible for alcoholic hepatosteatosis and ALD. The mechanism of ethanol action involves thedepletion of a unique RNA binding protein that specifically interacts with its 3'-UTR region of ST6Gal1 mRNA resulting in its destabilization and consequent appearance of asialoconjugates as alcohol biomarkers. With respect to ETOH effects on Cardio-Vascular Diseases, we conclude that CYP2E1 and ETOH mediated oxidative stress significantly down regulates not only the hepatic PON1 gene expression, but also serum PON1 and HCTLase activities accompanied by depletion of hepatic GSH, the endogenous antioxidant. These results strongly implicate the susceptibility of PON1 to increased ROS production. In contrast, betaine seems to be both hepatoprotective and atheroprotective by reducing hepatosteatosis and restoring not only liver GSH that quenches free radicals, but also the antiatherogenic PON1 gene expression and activity.

  12. CYP2E1 epigenetic regulation in chronic, low-level toluene exposure: Relationship with oxidative stress and smoking habit

    Energy Technology Data Exchange (ETDEWEB)

    Jiménez-Garza, Octavio, E-mail: ojimenezgarza@ugto.mx [Health Sciences Division, University of Guanajuato Campus León, Blvd. Puente del Milenio 1001, Fracción del Predio San Carlos, C.P. 37670 León, Guanajuato (Mexico); Baccarelli, Andrea A.; Byun, Hyang-Min [Laboratory of Environmental Epigenetics, Department of Environmental Health, Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, MA 02115 (United States); Márquez-Gamiño, Sergio [Health Sciences Division, University of Guanajuato Campus León, Blvd. Puente del Milenio 1001, Fracción del Predio San Carlos, C.P. 37670 León, Guanajuato (Mexico); Barrón-Vivanco, Briscia Socorro [Environmental Toxicology and Pollution Laboratory, Nayarit Autonomous University, Av. Ciudad de la Cultura s/n, “Amado Nervo”, Tepic, Nayarit C.P. 63155 (Mexico); Albores, Arnulfo [Department of Toxicology, CINVESTAV, Av. Instituto Politécnico Nacional 2508, Col. San Pedro Zacatenco, 07360 Mexico DF (Mexico)

    2015-08-01

    Background: CYP2E1 is a versatile phase I drug-metabolizing enzyme responsible for the biotransformation of most volatile organic compounds, including toluene. Human toluene exposure increases CYP2E1 mRNA and modifies its activity in leucocytes; however, epigenetic implications of this interaction have not been investigated. Goal: To determine promoter methylation of CYP2E1 and other genes known to be affected by toluene exposure. Methods: We obtained venous blood from 24 tannery workers exposed to toluene (mean levels: 10.86 +/− 7 mg/m{sup 3}) and 24 administrative workers (reference group, mean levels 0.21 +/− 0.02 mg/m{sup 3}) all of them from the city of León, Guanajuato, México. After DNA extraction and bisulfite treatment, we performed PCR-pyrosequencing in order to measure methylation levels at promoter region of 13 genes. Results: In exposed group we found significant correlations between toluene airborne levels and CYP2E1 promoter methylation (r = − .36, p < 0.05), as well as for IL6 promoter methylation levels (r = .44, p < 0.05). Moreover, CYP2E1 promoter methylation levels where higher in toluene-exposed smokers compared to nonsmokers (p = 0.009). We also observed significant correlations for CYP2E1 promoter methylation with GSTP1 and SOD1 promoter methylation levels (r = − .37, p < 0.05 and r = − .34, p < 0.05 respectively). Conclusion: These results highlight the importance of considering CYP2E1 epigenetic modifications, as well as its interactions with other genes, as key factors for unraveling the sub cellular mechanisms of toxicity exerted by oxidative stress, which can initiate disease process in chronic, low-level toluene exposure. People co-exposed to toluene and tobacco smoke are in higher risk due to a possible CYP2E1 repression. - Highlights: • We investigated gene-specific methylation in persons chronically exposed to toluene. • In a previous study, a reduced CYP2E1 activity was observed in these participants. • CYP2E1

  13. CYP2E1PstI/RsaI polymorphism and interaction with tobacco, alcohol and GSTs in gastric cancer susceptibility: A meta-analysis of the literature.

    NARCIS (Netherlands)

    S. Boccia (Stefania); S. de Lauretis (Angelo); F. Gianfagna (Francesco); C.M. van Duijn (Cock); G. Ricciardi (Gualtiero)

    2007-01-01

    textabstractStudies investigating the association between cytochrome P450 2E1 (CYP2E1) 5'-flanking region (PstI/RsaI) polymorphism and gastric cancer risk report conflicting results. The rationale for this meta-analysis was to determine whether CYP2E1*2 (c2) variant allele of CYP2E1 increases

  14. Effectiveness of Prostaglandin E1on hypoxic pulmonary hypertension in pig%前列腺素E1对缺氧性肺动脉高压的作用

    Institute of Scientific and Technical Information of China (English)

    孙依萍; 杨玲; 林建海

    2001-01-01

    目的 研究前列腺素E1在缺氧性肺动脉高压动物对血流动力学、血气分析和肺功能的作用。方法测定缺氧性肺动脉高压的小猪用前列腺素E1治疗前后的血流动力学、血气分析和肺功能的变化,在对照组测相同指标。结果 实验动物用前列腺素E1治疗后肺动脉压比治疗前显著下降(P<0.01),与对照组比较差异显著(P<0.01)。心搏指数在治疗后比治疗前增加(P<0.05),与对照组比较也有差异(P<0.05)。而血气分析、血压及肺功能无变化(P<0.05)。结论 前列腺素E1 能显著降低缺氧性肺动脉高压的肺动脉压,增加心输出量。%Objectives To investigate the effect of Prostagla n din E1on hemodynamic,blood gas analysis and lung function in animals with acu te hypoxic pulmonary hypertension.Methods Indices of hemodyna mic,blood gas analysis and lung function were detected before and after injectin g Prostaglandin E1in experimental pigs. While the same indices were detected in controls.Results Afer injecting intravenously Prostaglanden E1injection,the mean value of pulmonary artery pressure was significantly de creased than that before injecting Prostaglandin E1 in pigs and it was signif icantly decreased than that in controls(P<0.01);Cardiac Index was increase d in treatment group than that before injecting Prostaglandin E1 in pigs and it was increased than that in controls(P<0.05).Blood gas analysis,lung func tion were not different between groups(P>0.05).Conclusions The results suggested that Prostaglandin E1 could significantly decline the p ulmonary artery pressure and increase the cardiac output in animals with acute h ypoxic pulmonary hypertension.

  15. 细胞色素P4502E1的毒性机制研究进展%Research progress on toxicity mechanism of cytochrome P450 2E1

    Institute of Scientific and Technical Information of China (English)

    裴彦宇; 高虹

    2015-01-01

    细胞色素P4502E1(CYP2E1)在生物体内许多内源性或外源性物质的代谢过程中发挥着重要作用。CYP2E1主要分布在肝脏中,同时在肝外组织如心脏、肾脏等也有高表达。CYP2E1参与活性氧自由基的生成,引发氧化应激反应、脂质过氧化反应、炎性反应和细胞凋亡等过程,进而对机体产生毒性,诱发脂肪性肝病、糖尿病及肿瘤等疾病。本文综述了CYP2E1的毒性机制研究进展以及其在心脏毒性评价中的展望,其可能作为一个潜在的心脏毒性评价新指标应用于新药安全性评价中。%Cytochrome P450 2E1 (CYP2E1) in many endogenous or exogenous substances play an important role in the process of metabolism. CYP2E1 mainly expresses in liver, at the same time, in extra hepatic tissues such as heart, kid-ney and other place also has a high expression. CYP2E1 participates into the generate of reactive oxygen species, caus-es oxidant stress, lipid peroxidation, inflammatory response and apoptosis and so on, then produce toxicity to the body, and is related to fatty liver, diabetes, cancer, and so on. This article summarizes the toxicity mechanism of CYP2E1 re-search progress and the prospect of in the evaluation of cardiac toxicity, and may be as a potential new cardiac toxicity assessment indicators used in the safety evaluation of new drugs.

  16. Structure of Pyrazole Derivatives Impact their Affinity, Stoichiometry, and Cooperative Interactions for CYP2E1 Complexes

    Science.gov (United States)

    Hartman, Jessica H.; Bradley, Amber M.; Laddusaw, Ryan M; Perry, Martin D.; Miller, Grover P.

    2013-01-01

    CYP2E1 plays a critical role in detoxication and carcinogenic activation of drugs, pollutants, and dietary compounds; however, these metabolic processes can involve poorly characterized cooperative interactions that compromise the ability to understand and predict CYP2E1 metabolism. Herein, we employed an array of ten azoles with an emphasis on pyrazoles to establish the selectivity of catalytic and cooperative CYP2E1 sites through binding and catalytic studies. Spectral binding studies for monocyclic azoles suggested two binding events, while bicyclic azoles suggested one. Pyrazole had moderate affinity toward the CYP2E1 catalytic site that improved when a methyl group was introduced at either position 3 or 4. The presence of methyl groups simultaneously at positions 3 and 5 blocked binding, and a phenyl group at position 3 did not improve binding affinity. In contrast, pyrazole fusion to a benzene or cyclohexane ring greatly increased affinity. The consequences of these binding events on CYP2E1 catalysis were studied through inhibition studies with 4-nitrophenol, a substrate known to bind both sites. Most pyrazoles shared a common mixed cooperative inhibition mechanism in which pyrazole binding rescued CYP2E1 from substrate inhibition. Overall, inhibitor affinities toward the CYP2E1 catalytic site were similar to those reported in binding studies, and the same trend was observed for binding at the cooperative site. Taken together, these studies identified key structural determinants in the affinity and stoichiometry of azole interactions with CYP2E1 and consequences on catalysis that further advance an understanding of the relationship between structure and function for this enzyme. PMID:23811196

  17. pRB-E2F1 complexes are resistant to adenovirus E1A-mediated disruption.

    Science.gov (United States)

    Seifried, L A; Talluri, S; Cecchini, M; Julian, L M; Mymryk, J S; Dick, F A

    2008-05-01

    Disruption of pRB-E2F interactions by E1A is a key event in the adenoviral life cycle that drives expression of early viral transcription and induces cell cycle progression. This function of E1A is complicated by E2F1, an E2F family member that controls multiple processes besides proliferation, including apoptosis and DNA repair. Recently, a second interaction site in pRB that only contacts E2F1 has been discovered, allowing pRB to control proliferation separately from other E2F1-dependent activities. Based on this new insight into pRB-E2F1 regulation, we investigated how E1A affects control of E2F1 by pRB. Our data reveal that pRB-E2F1 interactions are resistant to E1A-mediated disruption. Using mutant forms of pRB that selectively force E2F1 to bind through only one of the two binding sites on pRB, we determined that E1A is unable to disrupt E2F1's unique interaction with pRB. Furthermore, analysis of pRB-E2F complexes during adenoviral infection reveals the selective maintenance of pRB-E2F1 interactions despite the presence of E1A. Our experiments also demonstrate that E2F1 functions to maintain cell viability in response to E1A expression. This suggests that adenovirus E1A's seemingly complex mechanism of disrupting pRB-E2F interactions provides selectivity in promoting viral transcription and cell cycle advancement, while maintaining cell viability.

  18. (E-1-Diphenylmethylidene-2-[(1H-indol-3-ylmethylidene]hydrazine

    Directory of Open Access Journals (Sweden)

    R. Archana

    2010-07-01

    Full Text Available In the title compound, C22H17N3, the 1H-indole unit is essentially planar, with a dihedral angle of 0.95 (10° between the pyrrole ring and the fused benzene ring. The dihedral angle between the two phenyl rings is 65.09 (10°. In the crystal, an intermolecular N—H...N hydrogen bond forms an infinite chain in the b-axis direction.

  19. (E)-1-Benzyl-idene-2,2-diphenyl-hydrazine.

    Science.gov (United States)

    Mendoza, Angel; Meléndrez-Luevano, Ruth; Cabrera-Vivas, Blanca M; Lozano-Márquez, Claudia D; Carranza, Vladimir

    2012-02-01

    The asymmetric unit of the title compound, C(19)H(16)N(2), contains two independent mol-ecules, both of which show an E configuration with respect to the C=N bond. The dihedral angles between the phenyl rings bonded to the hydrazine group are 81.00 (10) and 88.34 (8)° in the two mol-ecules. Inter-molecular C-H⋯π inter-actions are observed in the crystal structure.

  20. UNIT, TIBET.

    Science.gov (United States)

    Louisiana Arts and Science Center, Baton Rouge.

    THE UNIT OF STUDY DESCRIBED IN THIS BOOKLET DEALS WITH THE GEOGRAPHY AND HISTORY OF TIBET. THE UNIT COVERS SOME OF THE GENERAL FEATURES OF THE COUNTRY AND THEIR EFFECT UPON THE LIVES OF THE TIBETAN PEOPLE. DISCUSSION QUESTIONS ARE INSERTED TO STIMULATE THOUGHT. THE RELIGION OF TIBET IS DISCUSSED IN RELATION TO ITS INFLUENCE ON THE ART AND CULTURE…

  1. Low doses of nicotine and ethanol induce CYP2E1 and chlorzoxazone metabolism in rat liver.

    Science.gov (United States)

    Howard, L A; Micu, A L; Sellers, E M; Tyndale, R F

    2001-11-01

    The use of ethanol and nicotine is strongly linked; 80 to 95% of heavy alcohol users are also smokers. In humans, cigarette smoking significantly enhances CYP2E1 activity, as measured by increased metabolism of chlorzoxazone in vivo. CYP2E1 metabolizes ethanol and can generate toxic intermediates. CYP2E1 also bioactivates tobacco smoke and other procarcinogens and several hepatotoxins. We hypothesized that, like ethanol, nicotine increases CYP2E1 activity. Rats were treated once daily with saline, ethanol (0.3, 1.0, and 3.0 g/kg p.o.), or nicotine bitartrate (0.1, 0.3, and 1.0 mg base/kg s.c.) for 7 days. After ethanol or nicotine administration, immunostaining for CYP2E1 was increased in the centrilobular regions of rat liver. Western blot analyses revealed that hepatic CYP2E1 levels were increased by ethanol (1.6-2.4-fold) and nicotine (1.3-1.7-fold). In vitro chlorzoxazone 6-hydroxylation analyses demonstrated elevated Vmax values (compared with saline-treated animals) by using hepatic microsomes from high-dose ethanol (2.27 +/- 0.12 versus 1.18 +/- 0.23 nmol/mg/min, p neuropsychiatric motor disorders).

  2. In Silico Prediction of Human Sulfotransferase 1E1 Activity Guided by Pharmacophores from Molecular Dynamics Simulations.

    Science.gov (United States)

    Rakers, Christin; Schumacher, Fabian; Meinl, Walter; Glatt, Hansruedi; Kleuser, Burkhard; Wolber, Gerhard

    2016-01-01

    Acting during phase II metabolism, sulfotransferases (SULTs) serve detoxification by transforming a broad spectrum of compounds from pharmaceutical, nutritional, or environmental sources into more easily excretable metabolites. However, SULT activity has also been shown to promote formation of reactive metabolites that may have genotoxic effects. SULT subtype 1E1 (SULT1E1) was identified as a key player in estrogen homeostasis, which is involved in many physiological processes and the pathogenesis of breast and endometrial cancer. The development of an in silico prediction model for SULT1E1 ligands would therefore support the development of metabolically inert drugs and help to assess health risks related to hormonal imbalances. Here, we report on a novel approach to develop a model that enables prediction of substrates and inhibitors of SULT1E1. Molecular dynamics simulations were performed to investigate enzyme flexibility and sample protein conformations. Pharmacophores were developed that served as a cornerstone of the model, and machine learning techniques were applied for prediction refinement. The prediction model was used to screen the DrugBank (a database of experimental and approved drugs): 28% of the predicted hits were reported in literature as ligands of SULT1E1. From the remaining hits, a selection of nine molecules was subjected to biochemical assay validation and experimental results were in accordance with the in silico prediction of SULT1E1 inhibitors and substrates, thus affirming our prediction hypotheses.

  3. CYP2E1 epigenetic regulation in chronic, low-level toluene exposure: Relationship with oxidative stress and smoking habit.

    Science.gov (United States)

    Jiménez-Garza, Octavio; Baccarelli, Andrea A; Byun, Hyang-Min; Márquez-Gamiño, Sergio; Barrón-Vivanco, Briscia Socorro; Albores, Arnulfo

    2015-08-01

    CYP2E1 is a versatile phase I drug-metabolizing enzyme responsible for the biotransformation of most volatile organic compounds, including toluene. Human toluene exposure increases CYP2E1 mRNA and modifies its activity in leucocytes; however, epigenetic implications of this interaction have not been investigated. To determine promoter methylation of CYP2E1 and other genes known to be affected by toluene exposure. We obtained venous blood from 24 tannery workers exposed to toluene (mean levels: 10.86+/-7mg/m(3)) and 24 administrative workers (reference group, mean levels 0.21+/-0.02mg/m(3)) all of them from the city of León, Guanajuato, México. After DNA extraction and bisulfite treatment, we performed PCR-pyrosequencing in order to measure methylation levels at promoter region of 13 genes. In exposed group we found significant correlations between toluene airborne levels and CYP2E1 promoter methylation (r=-.36, ptoluene-exposed smokers compared to nonsmokers (p=0.009). We also observed significant correlations for CYP2E1 promoter methylation with GSTP1 and SOD1 promoter methylation levels (r=-.37, ptoluene exposure. People co-exposed to toluene and tobacco smoke are in higher risk due to a possible CYP2E1 repression. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Competitive Inhibition of Lysine Acetyltransferase 2B by a Small Motif of the Adenoviral Oncoprotein E1A.

    Science.gov (United States)

    Shi, Shasha; Liu, Ke; Chen, Yanheng; Zhang, Shijun; Lin, Juanyu; Gong, Chenfang; Jin, Quanwen; Yang, Xiang-Jiao; Chen, Ruichuan; Ji, Zhiliang; Han, Aidong

    2016-07-01

    The adenovirus early region 1A (E1A) oncoprotein hijacks host cells via direct interactions with many key cellular proteins, such as KAT2B, also known as PCAF (p300/CBP associated factor). E1A binds the histone acetyltransferase (HAT) domain of KAT2B to repress its transcriptional activation. However, the molecular mechanism by which E1A inhibits the HAT activity is not known. Here we demonstrate that a short and relatively conserved N-terminal motif (cNM) in the intrinsically disordered E1A protein is crucial for KAT2B interaction, and inhibits its HAT activity through a direct competition with acetyl-CoA, but not its substrate histone H3. Molecular modeling together with a series of mutagenesis experiments suggests that the major helix of E1A cNM binds to a surface of the acetyl-CoA pocket of the KAT2B HAT domain. Moreover, transient expression of the cNM peptide is sufficient to inhibit KAT2B-specific H3 acetylation H3K14ac in vivo Together, our data define an essential motif cNM in N-terminal E1A as an acetyl-CoA entry blocker that directly associates with the entrance of acetyl-CoA binding pocket to block the HAT domain access to its cofactor.

  5. Regulation of E1AF on matrix metaloproteinase-9 in colorectal cancer%大肠癌E1AF基因的表达对MMP-9的调控作用

    Institute of Scientific and Technical Information of China (English)

    朱民; 朱军

    2005-01-01

    目的:探讨大肠癌患者血浆基质金属蛋白酶-9(MMP-9)和肿瘤组织中E1A因子(E1AF)表达的临床意义及两者之间的关系.方法:采用酶链免疫吸附法(ELISA)检测51例大肠癌患者血浆MMP-9浓度,采用RT-PCR方法检测肿瘤组织中E1AF的表达,进行定量和半定量分析.结果:Ⅲ、Ⅳ期大肠癌患者血浆MMP-9浓度和肿瘤组织中E1AF的表达水平高于Ⅰ、Ⅱ期患者(P<0.05),Ⅳ期大肠癌患者MMP-9的浓度高于Ⅲ期患者(P<0.05);血浆MMP-9浓度和肿瘤组织中E1AF的表达呈正相关(r=0.77,P<0.05).结论:患者血浆MMP-9和肿瘤组织中E1AF的高表达与大肠癌浸润转移有关,E1AF参与MMP-9的表达调控.

  6. THE EFFECT OF PROSTAGLANDIN E1 ON PULMONARYBLOOD FLOW AFTER RETROGRADE FLUSH ANDCOLD STORAGE OF LUNGS

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Objective To determine the effects of prostaglandin E1 (PGE1) on the pulmonary blood flow by using re trograde flush (RF) in lung preservation. Methods Twelve canine donor lungs were treated with RF with UW solution. In group A (n=6), 250μg of PGE1 was infused into the pulmonary artery (PA) before RF and another 250μg was mixed with UW solution. No PGE1 was used in group B (n=6). After storage in the same solution at 4℃ for 22~25h, the left lung was transplanted to a recipient followed by 120min reperfusion. Pulmonary blood flow was determined by a microsphere technique. Colored microspheres were infused into the right atrium at the following time: before RF and donor lungs procurement (red), prior to resecting the recipient left lung (red, served as baseline), and 120min after donor lung implantation (yellow). Micro sphere distribution was expressed by optical density per gram lung tissue (OD/g). Results The red microsphere distribution after transplantation in group A was reduced as compared with baseline (P<0.05). And more blood flew to the right lung (P <0.05 ). The yellow microsphere distribution was also significantly reduced in group A when compared with that of group B (P <0.01 ). The hemodynamic data and blood gases for the two groups were not significantly different. Conclusion After single left lung transplantation, more blood flows to the right side. Pretreatment with PGE1 further reduces the flow distribution in the left side.

  7. Prostaglandin E1 inhibits collagenase gene expression in rabbit synoviocytes and human fibroblasts.

    Science.gov (United States)

    Salvatori, R; Guidon, P T; Rapuano, B E; Bockman, R S

    1992-07-01

    Cartilage breakdown, as seen in inflammatory and degenerative joint diseases, can be mediated by proteolytic enzymes, such as the metalloproteinase collagenase, the only enzyme able to digest collagen at neutral pH. In vitro collagenase gene expression can be stimulated by the phorbol ester tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate. We have investigated the effect of prostaglandin E1 (PGE1) on 12-O-tetradecanoyl-phorbol-13-acetate-stimulated collagenase mRNA levels in the rabbit synoviocyte cell line HIG-82. PGE1, but not PGE2 or PGF2 alpha, was able to selectively reduce collagenase mRNA levels in a dose-dependent fashion. PGE1 markedly increased intracellular levels of cAMP, while PGE2 and PGF2 alpha had little or no effect on cAMP production in the HIG-82 synoviocytes. Agents known to increase intracellular cAMP levels, such as the adenyl cyclase activator forskolin and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX), mimicked the effect of PGE1, on collagenase mRNA levels. PGE1, forskolin, and IBMX also decreased collagenase mRNA levels in human skin fibroblasts, demonstrating that this observation was not unique to the HIG-82 cell line. Transient transfection experiments carried out in HIG-82 cells using a 1.2-kilobase portion of the 5'-flanking region of the human collagenase gene linked to the reporter gene luciferase demonstrated that PGE1, forskolin, and IBMX exert their inhibitory effect on the promoter region of the collagenase gene.

  8. Combined prostaglandin E1 and lithium exert potent neuroprotection in a rat model of cerebral ischemia

    Institute of Scientific and Technical Information of China (English)

    Rui SHENG; Li-sha ZHANG; Rong HAN; Bo GAO; Xiao-qian LIU; Zheng-hong QIN

    2011-01-01

    Alm: To examine the effects of a mixed formulation composed of prostaglandin E1 and lithium (PGE1+Li mixture) on brain damage after cerebral ischemia. The effects of the mixture on protein expression of heat shock proteins (HSPs), p53, and Bcl-2 were also determined.Methods: Brain ischemia was induced with a permanent middle cerebral artery occlusion (pMCAO) in rats. Rats were treated with a single intravenous administration of PGE1, lithium or a PGE1+Li mixture immediately after the ischemic insult. The infarct volume and motor behavior deficits were analyzed 24 h after the ischemic insult. The protein levels of HSP70, glucose-regulated protein 78 (GRP78), HSP60, Bcl-2, and p53 in the striatum of the ipsilateral hemisphere were examined using immunoblotting.Results: The mixture (PGE1 22.6 nmol/kg+Li 0.5 mmol/kg) reduced infarct volume and neurological deficits induced by focal cerebral ischemia. Moreover, the mixture had a greater neuroprotective effect against cerebral ischemia compared with PGE1 or lithium alone.The mixture was effective even if it was administered 3 h after ischemia. PGE1+Li also significantly upregulated cytoprotective HSP70,GRP78, HSP60, and Bcl-2 protein levels, while decreasing p53 expression.Conclusion: These results demonstrated a PGE1+Li mixture with a therapeutic window of up to 3 h for clinical treatment of cerebral ischemia. The PGE1+Li mixture potentially exerts a protective effect after stroke through the induction of HSPs and Bcl-2 proteins.

  9. Inhibitory effects of prostaglandin E1 on activation of hepatic stellate cells in rabbits with schistosomiasis

    Institute of Scientific and Technical Information of China (English)

    Wei-Long Zou; Zhen Yang; Yun-Jin Zang; Dong-Jian Li; Zhi-Peng Liang; Zhong-Yang Shen

    2007-01-01

    BACKGROUND:Liver ifbrosis is the result of an imbalance between synthesis and degradation of extracellular matrix proteins of the liver. At the cellular and molecular levels, this progressive process is mainly characterized by activation of hepatic stellate cells (HSCs). Schistosoma japonica is one of the most prevalent causes of liver ifbrosis in China. It is characterized by hepatocyte damage, inlfammation, and chronic parasite egg-induced granuloma formation leading to ifbrosis. This study aimed to investigate the inhibitory effects of prostaglandin E1 (PGE1) on activation of HSCs and the alteration of type Ⅰ and Ⅲ collagen in rabbits with schistosomiasis. The study may promote the clinical application of praziquantel and PGE1 as a combined therapy to reverse hepatic ifbrosis caused by schistosomiasis. METHODS: Rabbits were percutaneously infected with cercaria of S. japonicum. Seven rabbits were subjected to intravenous injections of PGE1 (2.5 μg/kg daily) from days 60 to 120 after infection. The ultrastructural changes in activated HSCs were observed under transmission electron microscopy. The expression ofα-smooth muscle actin (α-SMA) was detected by immunohistochemistry. Fibril-forming collagens were detected by picrosirius staining. RESULTS: Activation of HSCs was a characteristic alteration in schistosome-induced hepatic ifbrosis. The expression of contraction-related α-SMA and the content of collagens were increased. Exogenous PGE1 markedly inhibited the activation of HSCs and reduced the expression of α-SMA around the hepatic sinusoids (P CONCLUSIONS:Activation of HSCs may play a key role in the progress of schistosome-induced hepatic ifbrosis. PGE1 effectively protects rabbit liver from ifbrosis, at least in part by inhibiting the activation of HSCs.

  10. Inositol hexakisphosphate inhibits mineralization of MC3T3-E1 osteoblast cultures.

    Science.gov (United States)

    Addison, William N; McKee, Marc D

    2010-04-01

    Inositol hexakisphosphate (IP6, phytic acid) is an endogenous compound present in mammalian cells and tissues. Differentially phosphorylated forms of inositol are well-documented to have important roles in signal transduction, cell proliferation and differentiation, and IP6 in particular has been suggested to inhibit soft tissue calcification (specifically renal and vascular calcification) by binding extracellularly to calcium oxalate and calcium phosphate crystals. However, the effects of IP6 on bone mineralization are largely unknown. In this study, we used MC3T3-E1 osteoblast cultures to examine the effects of exogenous IP6 on osteoblast function and matrix mineralization. IP6 at physiologic concentrations caused a dose-dependent inhibition of mineralization without affecting cell viability, proliferation or collagen deposition. Osteoblast differentiation markers, including tissue-nonspecific alkaline phosphatase activity, bone sialoprotein and osteocalcin mRNA levels, were not adversely affected by IP6 treatment. On the other hand, IP6 markedly increased protein and mRNA levels of osteopontin, a potent inhibitor of crystal growth and matrix mineralization. Inositol alone (without phosphate), as well as inositol hexakis-sulphate, a compound with a high negative charge similar to IP6, had no effect on mineralization or osteopontin induction. Binding of IP6 to mineral crystals from the osteoblast cultures, as well as to synthetic hydroxyapatite crystals, was confirmed by a colorimetric assay for IP6. In summary, IP6 inhibits mineralization of osteoblast cultures by binding to growing crystals through negatively charged phosphate groups and by induction of inhibitory osteopontin expression. These data suggest that IP6 may regulate physiologic bone mineralization by directly acting extracellularly, and by serving as a specific signal at the cellular level for the regulation of osteopontin gene expression.

  11. Crystal structures of (2E)-1-(3-bromo-thio-phen-2-yl)-3-(2-meth-oxy-phen-yl)prop-2-en-1-one and (2E)-1-(3-bromo-thio-phen-2-yl)-3-(3,4-di-meth-oxy-phen-yl)prop-2-en-1-one.

    Science.gov (United States)

    Naik, Vasant S; Shettigar, Venkataraya; Berglin, Tyler S; Coburn, Jillian S; Jasinski, Jerry P; Yathirajan, Hemmige S

    2015-08-01

    In the mol-ecules of the title compounds, (2E)-1-(3-bromo-thio-phen-2-yl)-3-(2-meth-oxy-phen-yl)prop-2-en-1-one, C14H11BrO2S, (I), which crystallizes in the space group P-1 with four independent mol-ecules in the asymmetric unit (Z' = 8), and (2E)-1-(3-bromo-thio-phen-2-yl)-3-(3,4-di-meth-oxy-phen-yl)prop-2-en-1-one, C15H13BrO3S, (II), which crystallizes with Z' = 8 in the space group I2/a, the non-H atoms are nearly coplanar. The mol-ecules of (I) pack with inversion symmetry stacked diagonally along the a-axis direction. Weak C-H⋯Br intra-molecular inter-actions in each of the four mol-ecules in the asymmetric unit are observed. In (II), weak C-H⋯O, bifurcated three-center inter-molecular inter-actions forming dimers along with weak C-H⋯π and π-π stacking inter-actions are observed, linking the mol-ecules into sheets along [001]. A weak C-H⋯Br intra-molecular inter-action is also present. There are no classical hydrogen bonds present in either structure.

  12. Role of cytochrome P450 2E1 in the metabolism of acrylamide and acrylonitrile in mice.

    Science.gov (United States)

    Sumner, S C; Fennell, T R; Moore, T A; Chanas, B; Gonzalez, F; Ghanayem, B I

    1999-11-01

    Acrylonitrile (AN) and acrylamide (AM) are commonly used in the synthesis of plastics and polymers. In rodents, AM and AN are metabolized to the epoxides glycidamide and cyanoethylene oxide, respectively. The aim of this study was to determine the role of cytochrome P450 in the metabolism of AM and AN in vivo. Wild-type (WT) mice, WT mice pretreated with aminobenzotriazole (ABT, 50 mg/kg ip, 2 h pre-exposure), and mice devoid of cytochrome P450 2E1 (P450 2E1-null) were treated with 50 mg/kg [(13)C]AM po. WT mice and P450 2E1-null mice were treated with 2.5 or 10 mg/kg [(13)C]AN po. Urine was collected for 24 h, and metabolites were characterized using (13)C NMR. WT mice excreted metabolites derived from the epoxides and from direct GSH conjugation with AM or AN. Only metabolites derived from direct GSH conjugation with AM or AN were observed in the urine from ABT-pretreated WT mice and P450 2E1-null mice. On the basis of evaluation of urinary metabolites at these doses, these data suggest that P450 2E1 is possibly the only cytochrome P450 enzyme involved in the metabolism of AM and AN in mice, that inhibiting total P450 activity does not result in new pathways of non-P450 metabolism of AM, and that mice devoid of P450 2E1 do not excrete metabolites of AM or AN that would be produced by oxidation by other cytochrome P450s. P450 2E1-null mice may be an appropriate model for the investigation of the role of oxidative metabolism in the toxicity or carcinogenicity of these compounds.

  13. A non-allergenic Ole e 1-like protein from birch pollen as a tool to design hypoallergenic vaccine candidates.

    Science.gov (United States)

    Marazuela, Eva G; Hajek, Roswitha; Villalba, Mayte; Barber, Domingo; Breiteneder, Heimo; Rodríguez, Rosalía; Batanero, Eva

    2012-02-01

    Recombinant DNA technology offers several approaches to convert allergens into hypoallergenic derivatives that can represent the basis of novel, safer and more effective forms of allergy vaccines. In this context, we used a new strategy for the design of a hypoallergenic derivative of Ole e 1, the main allergen of olive pollen. By screening a cDNA library from birch pollen, the clone BB18, encoding the birch counterpart of Ole e 1, was identified. In this study, BB18 has been produce in Pichia pastoris as a recombinant protein and immunologically characterized. The well-established non-allergenic properties of BB18 were used to generate a genetic variant of Ole e 1, named OB(55-58), by site-direct mutagenesis of four residues (E(55)V(56)G(57)Y(58)) in an IgE/IgG epitope of Ole e 1 by the corresponding ones in BB18 (SDSE). OB(55-58) was expressed in P. pastoris, purified to homogeneity and analyzed for IgE-reactivity by means of ELISA using sera from olive pollen allergic patients and rat basophil activation assay. T cell reactivity was assayed in a mouse model of Ole e 1 sensitization. The mutant OB(55-58) exhibited an impaired IgE reactivity, but not affected T cell reactivity, compared to wild type rOle e 1. This study emphasizes the usefulness of BB18 as a tool for epitope mapping and for engineering hypoallergenic derivatives of Ole e 1 as vaccine candidates for allergy prevention and treatment.

  14. Induction of brain cytochrome P450 2E1 boosts the locomotor-stimulating effects of ethanol in mice.

    Science.gov (United States)

    Ledesma, Juan Carlos; Miquel, Marta; Pascual, María; Guerri, Consuelo; Aragon, Carlos M G

    2014-10-01

    In the central nervous system ethanol (EtOH) is metabolized into acetaldehyde by different enzymes. Brain catalase accounts for 60% of the total production of EtOH-derived acetaldehyde, whereas cerebral cytochrome P450 2E1 (CYP 2E1) produces 20% of this metabolite. Acetaldehyde formed by the activity of central catalase has been implicated in some of the neurobehavioral properties of EtOH, yet the contribution of CYP 2E1 to the pharmacological actions of this drug has not been investigated. Here we assessed the possible participation of CYP 2E1 in the behavioral effects of EtOH. Thus, we induced CYP 2E1 activity and expression by exposing mice to chronic acetone intake (1% v/v for 10 days) and examined its consequences on the stimulating and uncoordinating effects of EtOH (0-3.2 g/kg) injected intraperitoneally. Our data showed that 24 h after withdrawal of acetone brain expression and activity of CYP 2E1 was induced. Furthermore, the locomotion produced by EtOH was boosted over the same interval of time. Locomotor stimulation produced by amphetamine or tert-butanol was unchanged by previous treatment with acetone. EtOH-induced motor impairment as evaluated in a Rota-Rod apparatus was unaffected by the preceding exposure to acetone. These results indicate that cerebral CYP 2E1 activity could contribute to the locomotor-stimulating effects of EtOH, and therefore we suggest that centrally produced acetaldehyde might be a possible mediator of some EtOH-induced pharmacological effects.

  15. Hepatic Lipid Peroxidation and Cytochrome P-450 2E1 in Pediatric Nonalcoholic Fatty Liver Disease and its Subtypes

    Science.gov (United States)

    Bell, Lauren N; Molleston, Jean P; Morton, Michael J; Klipsch, Ann; Saxena, Romil; Vuppalanchi, Raj; Chalasani, Naga

    2017-01-01

    Background Elevated hepatic oxidative stress and lipid peroxidation levels caused by increased hepatic Cytochrome P-450 2E1 (CYP2E1) enzyme activity has been speculated to play a role in the pathogenesis of nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH). But studies of lipid peroxidation and CYP2E1 in children are lacking. Aim To compare hepatic lipid peroxidation and hepatic CYP2E1 protein content in liver biopsies from children with NAFLD and two control groups. Study Liver biopsies from 59 children with NAFLD (49 with NASH), 10 children with normal liver histology, and 9 children with chronic hepatitis C infection (HCV) were examined. Hepatic malondialdehyde (MDA, a measure of lipid peroxidation) levels and CYP2E1 protein content were quantitated, as a percent of total area, by immunohistochemical staining of liver biopsy material followed by digital image quantitation. Results Lipid peroxidation was significantly greater in NAFLD liver biopsies (46.7 ± 20.8%) compared to liver biopsies from children with normal liver histology (7.6 ± 9.4%; pbiopsies compared to other groups (p=). Among children with NAFLD, lipid peroxidation and CYP2E1 protein content were not different between biopsies with and without NASH. The BMI was independently associated with hepatic lipid peroxidation (r=0.549; p<0.001). Conclusions Hepatic lipid peroxidation is increased in children with NAFLD but this is not related to hepatic CYP2E1. Lack of difference in lipid peroxidation among different NAFLD subgroups argues against its role in the disease progression. PMID:21602702

  16. Turnley Unit

    Data.gov (United States)

    Federal Laboratory Consortium — Facilities at this unit include cattle working pens, hydraulic squeeze chute and electronic scale, a maintenance building, and four hay storage sheds. There is one...

  17. Operable Units

    Data.gov (United States)

    U.S. Environmental Protection Agency — This dataset consists of operable unit data from multiple Superfund sites in U.S. EPA Region 8. These data were acquired from multiple sources at different times and...

  18. Detector Unit

    CERN Multimedia

    1960-01-01

    Original detector unit of the Instituut voor Kernfysisch Onderzoek (IKO) BOL project. This detector unit shows that silicon detectors for nuclear physics particle detection were already developed and in use in the 1960's in Amsterdam. Also the idea of putting 'strips' onto the silicon for high spatial resolution of a particle's impact on the detector were implemented in the BOL project which used 64 of these detector units. The IKO BOL project with its silicon particle detectors was designed, built and operated from 1965 to roughly 1977. Detector Unit of the BOL project: These detectors, notably the ‘checkerboard detector’, were developed during the years 1964-1968 in Amsterdam, The Netherlands, by the Natuurkundig Laboratorium of the N.V. Philips Gloeilampen Fabrieken. This was done in close collaboration with the Instituut voor Kernfysisch Onderzoek (IKO) where the read-out electronics for their use in the BOL Project was developed and produced.

  19. Absolute calibration of the Chang’E-1 IIM camera and its preliminary application

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    The interference imaging spectroradiometer (IIM) onboard the first lunar satellite of China "Chang’E-1" can now provide approximately global high spectral and spatial resolution reflectance spectra of the Moon. It is the essential instrument with which to accomplish one of the four missions of the first lunar satellite of China. As the current data provided by the Lunar Exploration Program Center and National Astronomical Observatories (NAOC) are not reflectance and the sensor response is inhomogeneous in the line direction,users can not use the current data directly. Moreover,due to the narrow band range,IIM data cannot cover the absorption peak of the mafic minerals of the Moon completely,which limits its ability for identifying minerals. The main objective of this study is to describe the methods for absolute calibration,correction and acquiring the absorption center of minerals for IIM data. The results from our study show that in the space domain the sensor response decreases toward the left,and in the spectral domain the response of the longer bands is more inhomogeneous than that of the shorter bands. After the calibration and correction,the reflectance of IIM matches the earth-based telescopic spectra well,which suggests the possible use of the processed data in the geological research. A high correlation was found between the absorption center and the wavelength at which the first derivative equals 0,i.e.,the so-called Stagnation Point in the mathematical sense. In the end,we show a preliminary applied study of the two craters with diameter larger than 35 km using the calibrated data. The spectra of IIM data show that the lunar crust has compositional diversity within the km scale. Pure anorthosite may be found on the wall and floor of the Aristarchus crater with the map of absorption center,which indicates that anorthosite is ubiquitously present within the lunar crust. IIM,with its capacity to acquire lunar composition at the regional and global scale

  20. Prostaglandin E1 alleviates neuropathic pain and neural dysfunction from entrapment neuropathy associated with diabetes mellitus.

    Science.gov (United States)

    Natsume, Tadahiro; Iwatsuki, Katsuyuki; Nishizuka, Takanobu; Arai, Tetsuya; Yamamoto, Michiro; Hirata, Hitoshi

    2014-10-01

    In this report, we present the results of investigation of the effects of prostaglandin E1 (PGE1) on entrapment neuropathy using a diabetic rat. A total of 60 male Sprague-Dawley rats were used in the study. The model of tibial nerve entrapment neuropathy associated with diabetes mellitus was created by streptozotocin-induced diabetic rats reared in cages with wire grid flooring. Rats were assigned to four groups: nondiabetic (n = 15), untreated diabetic (n = 15), diabetic treated with 30 μg/kg PGE1 (n = 15), and diabetic treated with 100 μg/kg PGE1 (n = 15). Pain tests and electrophysiological tests were performed at 0, 2, and 4 weeks, and assessments of gait, histology, and mRNA expression levels were performed at 4 weeks after initiating the PGE1 administration. In the 30 and 100 μg groups, the mechanical withdrawal thresholds measured by pain tests at 4 weeks (36.2 ± 16.4 g and 31.7 ± 15.3 g, respectively) and the motor conduction velocity (24.0 ± 0.2 m/s and 24.4 ± 0.3 m/s, respectively) were significantly higher than the untreated diabetic group (all P < 0.05) and lower than the nondiabetic group (all P < 0.001). In the gait analysis, the mean intensities in the 30 and 100 μg group (128.0 ± 20.1 a.u. and 109.0 ± 27.8 a.u., respectively) were significantly higher than the untreated diabetic (P < 0.01) and were not significantly different from the nondiabetic group (P = 0.81). Fiber density (P = 0.46) and fiber diameter (P = 0.15) did not show any significant differences. PGE1 significantly decreased nerve growth factor (NGF) mRNA and increased vascular endothelial growth factor (VEGF) mRNA in the tibial nerve (both P < 0.01). In conclusion, neurological deteriorations of diabetic rats were alleviated with PGE1, which is associated with inhibition of NGF and enhancement of VEGF at the entrapment site.

  1. Polyphosphates inhibit extracellular matrix mineralization in MC3T3-E1 osteoblast cultures.

    Science.gov (United States)

    Hoac, Betty; Kiffer-Moreira, Tina; Millán, José Luis; McKee, Marc D

    2013-04-01

    Studies on various compounds of inorganic phosphate, as well as on organic phosphate added by post-translational phosphorylation of proteins, all demonstrate a central role for phosphate in biomineralization processes. Inorganic polyphosphates are chains of orthophosphates linked by phosphoanhydride bonds that can be up to hundreds of orthophosphates in length. The role of polyphosphates in mammalian systems, where they are ubiquitous in cells, tissues and bodily fluids, and are at particularly high levels in osteoblasts, is not well understood. In cell-free systems, polyphosphates inhibit hydroxyapatite nucleation, crystal formation and growth, and solubility. In animal studies, polyphosphate injections inhibit induced ectopic calcification. While recent work has proposed an integrated view of polyphosphate function in bone, little experimental data for bone are available. Here we demonstrate in osteoblast cultures producing an abundant collagenous matrix that normally show robust mineralization, that two polyphosphates (PolyP5 and PolyP65, polyphosphates of 5 and 65 phosphate residues in length) are potent mineralization inhibitors. Twelve-day MC3T3-E1 osteoblast cultures with added ascorbic acid (for collagen matrix assembly) and β-glycerophosphate (a source of phosphate for mineralization) were treated with either PolyP5 or PolyP65. Von Kossa staining and calcium quantification revealed that mineralization was inhibited in a dose-dependent manner by both polyphosphates, with complete mineralization inhibition at 10μM. Cell proliferation and collagen assembly were unaffected by polyphosphate treatment, indicating that polyphosphate inhibition of mineralization results not from cell and matrix effects but from direct inhibition of mineralization. This was confirmed by showing that PolyP5 and PolyP65 bound to synthetic hydroxyapatite in a concentration-dependent manner. Tissue-nonspecific alkaline phosphatase (TNAP, ALPL) efficiently hydrolyzed the two PolyPs as

  2. The Deacetylase SIRT1 Regulates the Replication Properties of Human Papillomavirus 16 E1 and E2.

    Science.gov (United States)

    Das, Dipon; Smith, Nathan; Wang, Xu; Morgan, Iain M

    2017-05-15

    Human papillomaviruses (HPV) replicate their genomes in differentiating epithelium using the viral proteins E1 and E2 in association with host proteins. While the roles of E1 and E2 in this process are understood, the host factors involved and how they interact with and regulate E1-E2 are not. Our previous work identified the host replication and repair factor TopBP1 as an E2 partner protein essential for optimal E1-E2 replication and for the viral life cycle. The role of TopBP1 in host DNA replication is regulated by the class III deacetylase SIRT1; activation of the DNA damage response prevents SIRT1 deacetylation of TopBP1, resulting in a switch from DNA replication to repair functions for this protein and cell cycle arrest. Others have demonstrated an essential role for SIRT1 in regulation of the HPV31 life cycle; here, we report that SIRT1 can directly regulate HPV16 E1-E2-mediated DNA replication. SIRT1 is part of the E1-E2 DNA replication complex and is recruited to the viral origin of replication in an E1-E2-dependent manner. CRISPR/Cas9 was used to generate C33a clones with undetectable SIRT1 expression and lack of SIRT1 elevated E1-E2 DNA replication, in part due to increased acetylation and stabilization of the E2 protein in the absence of SIRT1. The results demonstrate that SIRT1 is a member of, and can regulate, the HPV16 replication complex. We discuss the potential role of this protein in the viral life cycle.IMPORTANCE HPV are causative agents in a number of human diseases, and currently only the symptoms of these diseases are treated. To identify novel therapeutic approaches for combating these diseases, the viral life cycle must be understood in more detail. This report demonstrates that a cellular enzyme, SIRT1, is part of the HPV16 DNA replication complex and is brought to the viral genome by the viral proteins E1 and E2. Using gene editing technology (CRISPR/Cas9), the SIRT1 gene was removed from cervical cancer cells. The consequence of this

  3. PDH-E1alpha dephosphorylation and activation in human skeletal muscle during exercise: effect of intralipid infusion.

    Science.gov (United States)

    Pilegaard, Henriette; Birk, Jesper B; Sacchetti, Massimo; Mourtzakis, Marina; Hardie, D Graham; Stewart, Greg; Neufer, P Darrell; Saltin, Bengt; van Hall, Gerrit; Wojtaszewski, Jorgen F P

    2006-11-01

    To investigate pyruvate dehydrogenase (PDH)-E1alpha subunit phosphorylation and whether free fatty acids (FFAs) regulate PDH activity, seven subjects completed two trials: saline (control) and intralipid/heparin (intralipid). Each infusion trial consisted of a 4-h rest followed by a 3-h two-legged knee extensor exercise at moderate intensity. During the 4-h resting period, activity of PDH in the active form (PDHa) did not change in either trial, yet phosphorylation of PDH-E1alpha site 1 (PDH-P1) and site 2 (PDH-P2) was elevated in the intralipid compared with the control trial. PDHa activity increased during exercise similarly in the two trials. After 3 h of exercise, PDHa activity remained elevated in the intralipid trial but returned to resting levels in the control trial. Accordingly, in both trials PDH-P1 and PDH-P2 decreased during exercise, and the decrease was more marked during intralipid infusion. Phosphorylation had returned to resting levels at 3 h of exercise only in the control trial. Thus, an inverse association between PDH-E1alpha phosphorylation and PDHa activity exists. Short-term elevation in plasma FFA at rest increases PDH-E1alpha phosphorylation, but exercise overrules this effect of FFA on PDH-E1alpha phosphorylation leading to even greater dephosphorylation during exercise with intralipid infusion than with saline.

  4. Garlic extract diallyl sulfide (DAS activates nuclear receptor CAR to induce the Sult1e1 gene in mouse liver.

    Directory of Open Access Journals (Sweden)

    Tatsuya Sueyoshi

    Full Text Available Constituent chemicals in garlic extract are known to induce phase I and phase II enzymes in rodent livers. Here we have utilized Car(+/+ and Car(-/- mice to demonstrate that the nuclear xenobiotic receptor CAR regulated the induction of the estrogen sulfotransferase Sult1e1 gene by diallyl sulfide (DAS treatment in mouse liver. DAS treatment caused CAR accumulation in the nucleus, resulting in a remarkable increase of SULT1E1 mRNA (3,200 fold and protein in the livers of Car(+/+ females but not of Car(-/- female mice. DAS also induced other CAR-regulated genes such as Cyp2b10, Cyp3a11 and Gadd45β. Compared with the rapid increase of these mRNA levels, which began as early as 6 hours after DAS treatment, the levels of SULT1E1 mRNA began increasing after 24 hours. This slow response to DAS suggested that CAR required an additional factor to activate the Sult1e1 gene or that this activation was indirect. Despite the remarkable induction of SULT1E1, there was no decrease in the serum levels of endogenous E2 or increase of estrone sulfate while the clearance of exogenously administrated E2 was accelerated in DAS treated mice.

  5. Cytochrome CYP2E1 phenotyping and genotyping in the evaluation of health risks from exposure to polluted environments.

    Science.gov (United States)

    Lucas, D; Ferrara, R; Gonzales, E; Albores, A; Manno, M; Berthou, F

    2001-10-15

    Humans are exposed to over 70,000 man-made chemicals including drugs, food additives, herbicides, pesticides, and industrial agents. It is well established that environmental chemicals are the cause of numerous human diseases including cancer. In most cases, chemical carcinogens require metabolic activation, which is mainly achieved by P450s enzymes. CYP2E1 is of clinical relevance because it is inducible by ethanol, and it metabolizes many common organic solvents such as benzene, alcohols and halogenated solvents. Therefore, alteration in the level of CYP2E1 might influence the health effects of the environmental pollutants. This hypothesis needs to be validated by epidemiological studies and the objective of the "Biomed-2" project was to develop new tests to assess the individual metabolic capacity of workers exposed to volatile organic compounds in order to predict their occupational risk. In vivo chlorzoxazone 6-hydroxylation was validated as a non-invasive and selective test for the determination of liver CYP2E1 activity. Preliminary data in workers exposed to organic solvents indicated that chlorzoxazone metabolism may be a biomarker of occupational exposure to organic solvents. Other approaches, such as use of salicylate as catalytic probe or measurement of catalytic activity in lymphocytes, were not conclusive. Attempts to use CYP2E1 genotyping for estimating human risks from chemical exposure did not bring convincing data as genetic polymorphism of CYP2E1 could not be clearly related to its catalytic activity.

  6. CYP2E1 gene rs6413420 polymorphism was first found in the Bouyei ethnic group of China.

    Science.gov (United States)

    Liu, Wei; Zhou, Li; Wang, Hongju; Zheng, Bo; Wu, Desheng; Yang, Xifei; Liu, Jianjun

    2014-01-01

    China is a multinational country. The relationship between gene polymorphisms of xenobiotic metabolizing enzymes and national ethnicity has not previously investigated among Chinese people. The aim of this study was to investigate distributions of CYP1A1 and CYP2E1 gene polymorphisms in five ethnic groups of China. 829 blood samples were collected from five ethnic groups (Han, Shui, Miao, Zhuang, Bouyei). Taqman-MGB probe was used in Real-time PCR to test the gene polymorphisms of CYP1A1 (rs1048943 and rs4646903) and CYP2E1 (rs2031920 and rs6413420). We further validate the SNP genotyping results through DNA sequencing. The genotype distribution of all four SNPs was in accordance with Hardy-Weinberg equilibrium except the genotype distribution of rs4646903 in Han and Bouyei ethnic groups (p=0.013 and 0.0005, respectively). CYP2E1 gene rs6413420 polymorphism was first found in the Bouyei ethnic group in China. The results of DNA sequencing were entirely in line with the SNP genotyping assay. The CYP1A1 and CYP2E1 genetic polymorphisms were different in different ethnic groups in China. CYP2E1 gene rs6413420 polymorphism was first found in the Bouyei ethnic group of China.

  7. Induction of CYP1A1 and CYP2E1 in rat liver by histamine: binding and kinetic studies.

    Science.gov (United States)

    Dávila-Borja, Víctor M; Belmont, Javier A; Espinosa, J Javier; Moreno-Sánchez, Rafael; Albores, Arnulfo; Montero, Regina D

    2007-10-01

    Histamine (HA) may bind to cytochrome P450 (CYP450) in rat liver microsomes. The CYP450-HA complex seems to regulate some cellular processes such as proliferation. In the present work, it is shown that HA increases the activity and protein level of CYP1A1 and CYP2E1, in vivo. CYP1A1 is associated with polycyclic aromatic hydrocarbon-mediated carcinogenesis and CYP2E1 with liver damage by oxidative stress. Studies of enzyme kinetics and binding with rat liver microsomes and supersomes were carried out to determine whether HA is a substrate of CYP1A1 and/or CYP2E1. The lack of NADPH oxidation in the presence of HA showed that it is not a substrate for CYP1A1. Activity measurements using the O-dealkylation of ethoxyresorufin indicated that HA is a mixed-type inhibitor of CYP1A1 in both microsomes and supersomes. On the other hand, HA induced a significant NADPH oxidation catalyzed by CYP2E1 supersomes, strongly suggesting that HA is a substrate for this isoform. Furthermore, HA is consumed in the presence of CYP2E1-induced microsomes and supersomes, as determined by o-phtalaldehyde complexes with HA by HPLC. The present findings may contribute to understand better the physiological function of CYP450 in relation with inflammation and other physiological processes in which HA may have a relevant role.

  8. Early-onset diabetic E1-DN mice develop albuminuria and glomerular injury typical of diabetic nephropathy.

    Science.gov (United States)

    Hyvönen, Mervi E; Dumont, Vincent; Tienari, Jukka; Lehtonen, Eero; Ustinov, Jarkko; Havana, Marika; Jalanko, Hannu; Otonkoski, Timo; Miettinen, Päivi J; Lehtonen, Sanna

    2015-01-01

    The transgenic E1-DN mice express a kinase-negative epidermal growth factor receptor in their pancreatic islets and are diabetic from two weeks of age due to impaired postnatal growth of β-cell mass. Here, we characterize the development of hyperglycaemia-induced renal injury in the E1-DN mice. Homozygous mice showed increased albumin excretion rate (AER) at the age of 10 weeks; the albuminuria increased over time and correlated with blood glucose. Morphometric analysis of PAS-stained histological sections and electron microscopy images revealed mesangial expansion in homozygous E1-DN mice, and glomerular sclerosis was observed in the most hyperglycaemic mice. The albuminuric homozygous mice developed also other structural changes in the glomeruli, including thickening of the glomerular basement membrane and widening of podocyte foot processes that are typical for diabetic nephropathy. Increased apoptosis of podocytes was identified as one mechanism contributing to glomerular injury. In addition, nephrin expression was reduced in the podocytes of albuminuric homozygous E1-DN mice. Tubular changes included altered epithelial cell morphology and increased proliferation. In conclusion, hyperglycaemic E1-DN mice develop albuminuria and glomerular and tubular injury typical of human diabetic nephropathy and can serve as a new model to study the mechanisms leading to the development of diabetic nephropathy.

  9. Early-Onset Diabetic E1-DN Mice Develop Albuminuria and Glomerular Injury Typical of Diabetic Nephropathy

    Directory of Open Access Journals (Sweden)

    Mervi E. Hyvönen

    2015-01-01

    Full Text Available The transgenic E1-DN mice express a kinase-negative epidermal growth factor receptor in their pancreatic islets and are diabetic from two weeks of age due to impaired postnatal growth of β-cell mass. Here, we characterize the development of hyperglycaemia-induced renal injury in the E1-DN mice. Homozygous mice showed increased albumin excretion rate (AER at the age of 10 weeks; the albuminuria increased over time and correlated with blood glucose. Morphometric analysis of PAS-stained histological sections and electron microscopy images revealed mesangial expansion in homozygous E1-DN mice, and glomerular sclerosis was observed in the most hyperglycaemic mice. The albuminuric homozygous mice developed also other structural changes in the glomeruli, including thickening of the glomerular basement membrane and widening of podocyte foot processes that are typical for diabetic nephropathy. Increased apoptosis of podocytes was identified as one mechanism contributing to glomerular injury. In addition, nephrin expression was reduced in the podocytes of albuminuric homozygous E1-DN mice. Tubular changes included altered epithelial cell morphology and increased proliferation. In conclusion, hyperglycaemic E1-DN mice develop albuminuria and glomerular and tubular injury typical of human diabetic nephropathy and can serve as a new model to study the mechanisms leading to the development of diabetic nephropathy.

  10. Interaction of Adenovirus E1A with the HHV8 Promoter of Latent Genes: E1A Proteins are Able to Activate the HHV-8 LANAp in MV3 Reporter Cells

    OpenAIRE

    Koehler-Hansner, Karin; Flore, Ornella; Opalka, Bertram; Hengge, Ulrich R.

    2008-01-01

    Human herpesvirus 8 (HHV-8) is associated with Kaposi’s sarcoma, body cavity-based lymphoma, and Castleman’s disease. Adenoviral (Ad) E1A proteins regulate the activity of cellular and viral promoters/enhancers and transcription factors and can suppress tumorigenicity of human cancers. As (i) HHV-8 and Ad may co-exist in immunocompromised patients and (ii) E1A might be considered as therapeutic transgene for HHV-8-associated neoplasms we investigated whether the promoter of the latency-associ...

  11. A p53-independent apoptotic mechanism of adenoviral mutant E1A was involved in its selective antitumor activity for human cancer

    Science.gov (United States)

    Fang, Lin; Cheng, Qian; Zhao, Jingjing; Ge, Yan; Zhu, Qi; Zhao, Min; Zhang, Jie; Zhang, Qi; Li, Liantao; Liu, Junjie; Zheng, Junnian

    2016-01-01

    The conserved regions (CR) of adenoviral E1A had been shown to be necessary for disruption of pRb-E2F transcription factor complexes and induction of the S phase. Here we constructed a mutant adenoviral E1A with Rb-binding ability absent (E1A 30-60aa and 120-127aa deletion, mE1A) and investigated its antitumor capacities in vitro and in vivo. The mE1A suppressed the viability of tumor cells as efficiently as the wild type E1A, and there was no cytotoxic effect on normal cells. Although the mE1A arrested tumor cell cycle with the same manner as E1A, the former played a different role on cell cycle regulation compared with E1A in normal cells, which might contribute to its selective antitumor activity. E1A and mE1A had accumulated inactive p53, decreased the expression of mdm2, Cdkn1a (also named p21), increased p21's nuclear distribution and induced tumor cell apoptosis in a p53-indenpent manner. Further, E1A or mE1A significantly suppressed tumor growth in subcutaneous hepatocellular carcinoma xenograft models. Especially, tumor-bearing mice treated with mE1A had higher survival rate than those treated with E1A. Our data demonstrated that mutant adenoviral E1A significantly induced tumor cell apoptosis in a p53-indenpednt manner and had selective tumor suppressing ability. The observations of adenoviral E1A mutant had provided a novel mechanism for E1A's complex activities during infection. PMID:27340782

  12. (4E-1-Phenyl-4-{[(pyridin-2-ylamino]methylidene}pyrazolidine-3,5-dione

    Directory of Open Access Journals (Sweden)

    Shaaban K. Mohamed

    2016-06-01

    Full Text Available The title compound, C15H12N4O2, contains two independent molecules in the asymmetric unit. These differ in terms of dihedral angles that the phenyl and 2-pyridyl rings subtend with the central five-membered ring; 15.2 (2 and 2.9 (2°, respectively, in one molecule, 8.9 (2 and 5.1 (2°for the second. In the crystal, the independent molecules each self-associate to form layers through N—H...O and C—H...O hydrogen bonding. The layers associate through π–π interactions between the phenyl rings and isolated carbon–carbon double bonds [shortest midpoint–centroid distance = 3.347 (4 Å]. The crystal studied was refined as a two-component twin.

  13. (E-1-(2,4-Dimethylquinolin-3-yl-3-(4-methylphenylprop-2-en-1-one

    Directory of Open Access Journals (Sweden)

    R. Prasath

    2012-05-01

    Full Text Available In the title compound, C21H19NO, there are two molecules in the asymmetric unit (Z′ = 2. There are π–π interactions between these two molecules [centroid–centroid distance = 3.678 (2 Å], as well as a weak C—H...O interaction. The conformation adopted by the two molecules is such that the quinoline mean plane and the benzene ring are almost perpendicular [89.04 (5 and 76.89 (4°]. In each molecule, the methyl group of the tolyl ring is disordered over two conformations, with occupancy ratios of 0.56 (3:0.44 (3 and 0.65 (3:0.35 (3.

  14. Tmed2 accelerates murine MC3T3-E1 pre-osteoblast proliferation in vitro%Tmed2促进体外小鼠前成骨细胞MC3T3-E1增殖

    Institute of Scientific and Technical Information of China (English)

    熊元; 熊霞辉; 卢雅彬; 朱宁; 陈梅红

    2012-01-01

    目的 研究Tmed2基因对小鼠前成骨细胞增殖的影响.方法1)分别在小鼠MC3T3-E1细胞中过表达和抑制Tmed2,检测细胞增殖情况.2)用雌激素处理细胞后,检测细胞的增殖及Tmed2基因的表达量.荧光实时定量PCR检测mRNA水平,MTS法检测细胞活力和增殖,流式细胞术检测细胞周期,Western blot法检测蛋白水平.结果 过表达Tmed2使MC3T3 -E1细胞的增殖速度加快,细胞周期中S期细胞比例明显增加,且Cyclin A的表达升高.而抑制Tmed2基因表达使MC3T3-E1细胞的增殖速度减慢.雌激素处理使细胞增殖速度加快的同时,Tmed2基因的表达显著增高.结论 Tmed2通过上调Cyclin A的表达,使S期细胞比例增加,加快小鼠前成骨细胞MC3T3-E1的增殖.此外,Tmed2的表达受雌激素的调控,可能参与雌激素促进MC3T3-E1细胞增殖的作用.%Objective To study the role of Tmed2 in murine pre-osteoblast proliferation. Methods 1) Over-express and knock down Tmed2, and the proliferation rate of MC3T3-E1 cell was then detected. 2) MC3T3-E1 cells were treated with p-Estradiol, cell number was then counted and the mRNA level of Tmed2 was analyzed. Quantitative real-time PCR was used to measure the mRNA level, MTS assay was used to assess the cell proliferation rate, flow eytometry was used to figure out the cell cycle distribution, and the protein level was evaluated by Western blot. Results Over-expression of Tmed2 accelerated MC3T3-E1 cell proliferation, and the proportion of cells in S phase markedly increased. The expression of Cyclin A also increased. On the contrary, Knockdown of Tmed2 decelerated MC3T3-E1 cell proliferation. In β-Estradiol promoted MC3T3-E1 proliferation, the mRNA level of Tmed2 remarkably increased. Conclusions Tmed2 accelerates murine pre-osteoblast MC3T3-E1 proliferation through up-regulating Cyclin A expression and therefore increasing the proportion of cells in S phase. In addition, the expression of Tmed2 is regulated by

  15. Painful Charcot-Marie-Tooth neuropathy type 2E/1F due to a novel NEFL mutation.

    Science.gov (United States)

    Doppler, Kathrin; Kunstmann, Erdmute; Krüger, Stefan; Sommer, Claudia

    2017-05-01

    Charcot-Marie-Tooth neuropathy (CMT) 2E/1F is caused by mutations in the neurofilament light-chain polypeptide (NEFL) gene. Giant axons are a histological hallmark frequently seen in nerves of patients with CMT2E. We describe the case of a 43-year-old patient with a painful, predominantly sensory neuropathy. The patient's sural nerve biopsy showed multiple giant axons. Genetic sequencing of the NEFL gene revealed that the patient was heterozygous for an altered sequence of the gene, c.816C>G, p.Asn272Lys, which has not yet been described in CMT2E/1F. In contrast to other cases of CMT2E/1F, where motor symptoms are predominant, pain was the most disabling symptom in this patient. Muscle Nerve 55: 752-755, 2017. © 2016 Wiley Periodicals, Inc.

  16. Green tea polyphenol (-)-epigallocatechin gallate suppressed the differentiation of murine osteoblastic MC3T3-E1 cells.

    Science.gov (United States)

    Kamon, Masayoshi; Zhao, Ran; Sakamoto, Kazuichi

    2009-12-16

    Recently, various physiological effects of the tea polyphenol catechin for alleviating diseases such as cancer, arteriosclerosis, hyperlipidaemia and osteoporosis have been reported. However, the physiological effect of catechin on bone metabolism remains unclear. We examined the physiological effect of EGCG [(-)-epigallocatechin-3-gallate], which is the main component of green tea catechin, on osteoblast development using the precursor cell line of osteoblasts, MC3T3-E1, and co-culture of the osteoblasts from mouse newborn calvaria and mouse bone marrow cells. Although EGCG did not affect the viability and proliferation of MC3T3-E1 cells, EGCG inhibited the osteoblast differentiation. Furthermore, EGCG did not affect the mineralization of differentiated MC3T3-E1 cells, and reduced osteoclast formation in co-culture. These results suggest that EGCG can effectively suppress bone resorption, and can be used as an effective medicine in the treatment of the symptoms of osteoporosis.

  17. Genotype and allele frequencies of polymorphic cytochromes P450 CYP1A2 and CYP2E1 in Mexicans.

    Science.gov (United States)

    Mendoza-Cantú, Ania; Castorena-Torres, Fabiola; Bermudez, Mario; Martínez-Hernández, Roberto; Ortega, Arturo; Salinas, Juan E; Albores, Arnulfo

    2004-01-01

    CYP1A2 and CYP2E1 are two of the main cytochrome P450 isoforms involved in the metabolism of commonly used drugs and xenobiotic compounds considered to be responsible for or possible participants in the development of several human diseases. Individual susceptibility to developing these pathologies relies, among other factors, on genetic polymorphism which depends on ethnic differences, as the frequency of mutant genotypes varies in different human populations. Thus the aim of this study was to investigate the frequency of CYP1A2 5'-flanking region and CYP2E1 Rsa I/Pst I polymorphisms in Mexicans by PCR-RFLP methods. The DNA of 159 subjects was analysed and mutant allele frequencies of 30% for CYP2E1 Rsa I/Pst I sites and 43% for CYP1A2 5'-flanking region were found. These frequencies are higher than those previously reported for other human populations.

  18. Unit Replacement System Analysis I (URSA I).

    Science.gov (United States)

    1982-01-01

    action creates a void in CONUS of one unit. A new unit is formed in CONUS to fill this void . The forming of a new unit is 3-5 -,.. - ..-- o. . CAA-SR-82...compares the observed average regimental strengths with the respective authorizations. Note the E-1 to E-4 and E-5 overfill and the E-6 to E-8 underfill ...When the data is adjusted to compensate for grade substitution flexibility (see Figure 6-26), the E-6 to E-8 underfill is corrected, but the

  19. Ash pollen allergy: reliable detection of sensitization on the basis of IgE to Ole e 1.

    Science.gov (United States)

    Imhof, Konrad; Probst, Elisabeth; Seifert, Burkhardt; Regenass, Stephan; Schmid-Grendelmeier, Peter

    Background: Alongside hazel, alder and birch pollen allergies, ash pollen allergy is a relevant cause of hay fever during spring in the European region. For some considerable time, ash pollen allergy was not routinely investigated and its clinical relevance may well have been underestimated, particularly since ash and birch tree pollination times are largely the same. Ash pollen extracts are not yet well standardized and diagnosis is therefore sometimes unreliable. Olive pollen, on the other hand, is strongly cross-reactive with ash pollen and is apparently better standardized. Therefore, the main allergen of olive pollen, Ole e 1, has been postulated as a reliable alternative for the detection of ash pollen sensitization. Methods: To determine to what extent specific IgE against Ole e 1 in patients with ash pollen allergy is relevant, we included 183 subjects with ash pollen allergy displaying typical symptoms in March/April and positive skin prick test specific IgE against Ole e 1 (t224) and ash pollen (t25) and various birch allergens (Bet v 1, Bet v 2/v 4) in a retrospective study. Results: A significant correlation was seen between specific IgE against Ole e 1 and ash pollen, but also to a slightly lesser extent between IgE against Ole e 1 and skin prick test with ash pollen, the latter being even higher than IgE and skin prick test both with ash pollen. No relevant correlation was found with birch pollen allergens, demonstrating the very limited cross-reactivity between ash and birch pollen. Conclusion: It appears appropriate to determine specific IgE against Ole e 1 instead of IgE against ash pollen to detect persons with ash pollen allergy. Our findings may also support the idea of using possibly better standardized or more widely available olive pollen extracts instead of ash pollen extract for allergen-specific immunotherapy.

  20. Occupational Toluene Exposure Induces Cytochrome P450 2E1 mRNA Expression in Peripheral Lymphocytes

    Science.gov (United States)

    Mendoza-Cantú, Ania; Castorena-Torres, Fabiola; de León, Mario Bermúdez; Cisneros, Bulmaro; López-Carrillo, Lizbeth; Rojas-García, Aurora E.; Aguilar-Salinas, Alberto; Manno, Maurizio; Albores, Arnulfo

    2006-01-01

    Print workers are exposed to organic solvents, of which the systemic toxicant toluene is a main component. Toluene induces expression of cytochrome P450 2E1 (CYP2E1), an enzyme involved in its own metabolism and that of other protoxicants, including some procarcinogens. Therefore, we investigated the association between toluene exposure and the CYP2E1 response, as assessed by mRNA content in peripheral lymphocytes or the 6-hydroxychlorzoxazone (6OH-CHZ)/chlorzoxazone (CHZ) quotient (known as CHZ metabolic ratio) in plasma, and the role of genotype (5′-flanking region RsaI/PstI polymorphic sites) in 97 male print workers. The geometric mean (GM) of toluene concentration in the air was 52.80 ppm (10–760 ppm); 54% of the study participants were exposed to toluene concentrations that exceeded the maximum permissible exposure level (MPEL). The GM of urinary hippuric acid at the end of a work shift (0.041 g/g creatinine) was elevated relative to that before the shift (0.027 g/g creatinine; p < 0.05). The GM of the CHZ metabolic ratio was 0.33 (0–9.3), with 40% of the subjects having ratios below the GM. However, the average CYP2E1 mRNA level in peripheral lymphocytes was 1.07 (0.30–3.08), and CYP2E1 mRNA levels within subjects correlated with the toluene exposure ratio (environmental toluene concentration:urinary hippuric acid concentration) (p = 0.014). Genotype did not alter the association between the toluene exposure ratio and mRNA content. In summary, with further validation, CYP2E1 mRNA content in peripheral lymphocytes could be a sensitive and noninvasive biomarker for the continuous monitoring of toluene effects in exposed persons. PMID:16581535

  1. Genotype and allelic frequencies of CYP2E1*5B polymorphism in the southwest population of Iran

    Directory of Open Access Journals (Sweden)

    Fatemeh Zanganeh

    2014-10-01

    Full Text Available Background: Cytochrome P450 2E1 (CYP2E1 is a main enzyme which plays a major role in activating and detoxifying many xenobiotics, carcinogens and drugs. Available studies suggest that CYP2E1 single nucleotide polymorphisms (SNPs are involved in the risk of developing certain cancers after exposure to carcinogens. The purpose of the present study was to assess genotype and allele frequencies of polymorphic CYP2E1*5B in the Iranian population. Material and Methods: This study was performed on 200 healthy individuals (female: 100, male: 100 in medical laboratories of Ahvaz during 2011. The CYP2E1 *5B (rs3813867 G-1293C assessment was carried out using PCR-RFLP method. The data were analyzed with ĸ2 and hardy-Weinberg Equation statistically methods. Results: The frequency of *1A/*1A (c1/c1, *1A/*5B (c1/c2 and *5B/*5B (c2/c2 genotypes was computed 97, 3 and 0 percent, respectively. The frequency of *1A (c1 and *5B (c2 alleles was computed 98.5 and 1.5 percent, respectively. No statistically significant difference was between two genders (p>0.05. Conclusion: The genotype distribution and allele frequencies of CYP2E1*5B polymorphism were similar to Turkish and some of the European populations. However, there are significant interethnic differences when the Iranian population is compared with the Eastern Asian, American and some of the European populations. The allelic distribution of this polymorphism did not vary with gender.

  2. Occupational toluene exposure induces cytochrome P450 2E1 mRNA expression in peripheral lymphocytes.

    Science.gov (United States)

    Mendoza-Cantú, Ania; Castorena-Torres, Fabiola; Bermúdez de León, Mario; Cisneros, Bulmaro; López-Carrillo, Lizbeth; Rojas-García, Aurora E; Aguilar-Salinas, Alberto; Manno, Maurizio; Albores, Arnulfo

    2006-04-01

    Print workers are exposed to organic solvents, of which the systemic toxicant toluene is a main component. Toluene induces expression of cytochrome P450 2E1 (CYP2E1), an enzyme involved in its own metabolism and that of other protoxicants, including some procarcinogens. Therefore, we investigated the association between toluene exposure and the CYP2E1 response, as assessed by mRNA content in peripheral lymphocytes or the 6-hydroxychlorzoxazone (6OH-CHZ)/chlorzoxazone (CHZ) quotient (known as CHZ metabolic ratio) in plasma, and the role of genotype (5 -flanking region RsaI/PstI polymorphic sites) in 97 male print workers. The geometric mean (GM) of toluene concentration in the air was 52.80 ppm (10-760 ppm); 54% of the study participants were exposed to toluene concentrations that exceeded the maximum permissible exposure level (MPEL). The GM of urinary hippuric acid at the end of a work shift (0.041 g/g creatinine) was elevated relative to that before the shift (0.027 g/g creatinine; p < 0.05). The GM of the CHZ metabolic ratio was 0.33 (0-9.3), with 40% of the subjects having ratios below the GM. However, the average CYP2E1 mRNA level in peripheral lymphocytes was 1.07 (0.30-3.08), and CYP2E1 mRNA levels within subjects correlated with the toluene exposure ratio (environmental toluene concentration:urinary hippuric acid concentration) (p = 0.014). Genotype did not alter the association between the toluene exposure ratio and mRNA content. In summary, with further validation, CYP2E1 mRNA content in peripheral lymphocytes could be a sensitive and noninvasive biomarker for the continuous monitoring of toluene effects in exposed persons.

  3. CYP2E1 regulation by benzene and other small organic chemicals in rat liver and peripheral lymphocytes.

    Science.gov (United States)

    González-Jasso, Eva; López, Tomás; Lucas, Daniele; Berthou, Francois; Manno, Maurizio; Ortega, Arturo; Albores, Arnulfo

    2003-09-15

    The inducibility of CYP2E1 was investigated in liver and peripheral lymphocytes of rats treated with benzene (0-10 mmol/kg body weight (bw), daily for 3 days, i.p., or 0 and 5 mmol/kg bw, daily for 14 days, i.p.) or toluene (0 and 5 mmol/kg bw, daily for 3 days, i.p.) and compared with that of pyridine (5 mmol/kg bw, i.p.) or acetone (5% in drinking water) both daily for 3 days. Acute benzene treatment (5 mmol/kg bw) increased both CYP2E1 apo-protein (2-fold) and p-nitrophenol hydroxylase (p-NPH) activity (1.4-fold) in liver, and CYP2E1 mRNA in both liver (2.2-fold) and peripheral lymphocytes (2.9-fold). The response to toluene was qualitatively similar, although smaller than that to benzene. As expected, acetone and pyridine treatments resulted in a 2- to 3-fold increase of p-NPH activity and CYP2E1 apo-protein content in liver, but not the mRNA levels. In addition, acute benzene and acetone treatments increased the 6-hydroxychlorzoxazone/chlorzoxazone metabolic ratio 1.6- and 3.1-fold, respectively. The subchronic treatment with benzene increased CYP2E1 mRNA and apo-protein from days 2 and 3 to day 14, respectively, whereas the enzyme activity increased transiently on days 3 and 5 only. These results show that acute/subacute benzene and acute toluene treatments induce CYP2E1 expression probably through a similar mechanism which might be different from that of pyridine or acetone, in that the former increase mRNA levels, both in liver and in peripheral lymphocytes, whereas the latter stabilized the apo-protein.

  4. Incomplete LPS Core-Specific Felix01-Like Virus vB_EcoM_VpaE1.

    Science.gov (United States)

    Šimoliūnas, Eugenijus; Vilkaitytė, Monika; Kaliniene, Laura; Zajančkauskaitė, Aurelija; Kaupinis, Algirdas; Staniulis, Juozas; Valius, Mindaugas; Meškys, Rolandas; Truncaitė, Lidija

    2015-11-27

    Bacteriophages represent a valuable source for studying the mechanisms underlying virus-host interactions. A better understanding of the host-specificity of viruses at the molecular level can promote various phage applications, including bacterial diagnostics, antimicrobial therapeutics, and improve methods in molecular biology. In this study, we describe the isolation and characterization of a novel coliphage, vB_EcoM_VpaE1, which has different host specificity than its relatives. Morphology studies, coupled with the results of genomic and proteomic analyses, indicate that vB_EcoM_VpaE1 belongs to the newly proposed genus Felix01likevirus in the family Myoviridae. The genus Felix01likevirus comprises a group of highly similar phages that infect O-antigen-expressing Salmonella and Escherichia coli (E. coli) strains. Phage vB_EcoM_VpaE1 differs from the rest of Felix01-like viruses, since it infects O-antigen-deficient E. coli strains with an incomplete core lipopolysaccharide (LPS). We show that vB_EcoM_VpaE1 can infect mutants of E. coli that contain various truncations in their LPS, and can even recognize LPS that is truncated down to the inner-core oligosaccharide, showing potential for the control of rough E. coli strains, which usually emerge as resistant mutants upon infection by O-Ag-specific phages. Furthermore, VpaE1 can replicate in a wide temperature range from 9 to 49 °C, suggesting that this virus is well adapted to harsh environmental conditions. Since the structural proteins of such phages tend to be rather robust, the receptor-recognizing proteins of VpaE1 are an attractive tool for application in glycan analysis, bacterial diagnostics and antimicrobial therapeutics.

  5. Incomplete LPS Core-Specific Felix01-Like Virus vB_EcoM_VpaE1

    Directory of Open Access Journals (Sweden)

    Eugenijus Šimoliūnas

    2015-11-01

    Full Text Available Bacteriophages represent a valuable source for studying the mechanisms underlying virus-host interactions. A better understanding of the host-specificity of viruses at the molecular level can promote various phage applications, including bacterial diagnostics, antimicrobial therapeutics, and improve methods in molecular biology. In this study, we describe the isolation and characterization of a novel coliphage, vB_EcoM_VpaE1, which has different host specificity than its relatives. Morphology studies, coupled with the results of genomic and proteomic analyses, indicate that vB_EcoM_VpaE1 belongs to the newly proposed genus Felix01likevirus in the family Myoviridae. The genus Felix01likevirus comprises a group of highly similar phages that infect O-antigen-expressing Salmonella and Escherichia coli (E. coli strains. Phage vB_EcoM_VpaE1 differs from the rest of Felix01-like viruses, since it infects O-antigen-deficient E. coli strains with an incomplete core lipopolysaccharide (LPS. We show that vB_EcoM_VpaE1 can infect mutants of E. coli that contain various truncations in their LPS, and can even recognize LPS that is truncated down to the inner-core oligosaccharide, showing potential for the control of rough E. coli strains, which usually emerge as resistant mutants upon infection by O-Ag-specific phages. Furthermore, VpaE1 can replicate in a wide temperature range from 9 to 49 °C, suggesting that this virus is well adapted to harsh environmental conditions. Since the structural proteins of such phages tend to be rather robust, the receptor-recognizing proteins of VpaE1 are an attractive tool for application in glycan analysis, bacterial diagnostics and antimicrobial therapeutics.

  6. Short-term treatment with alcohols causes hepatic steatosis and enhances acetaminophen hepatotoxicity in Cyp2e1(-/-) mice.

    Science.gov (United States)

    Sinclair, J F; Szakacs, J G; Wood, S G; Walton, H S; Bement, J L; Gonzalez, F J; Jeffery, E H; Wrighton, S A; Bement, W J; Sinclair, P R

    2000-10-15

    CYP2E1 has been reported to have an essential role in alcohol-mediated increases in hepatic steatosis and acetaminophen hepatotoxicity. We found that pretreatment of Cyp2e1(-/-) mice with ethanol plus isopentanol, the predominant alcohols in alcoholic beverages, for 7 days resulted in micro- and macrovesicular steatosis in the livers of all mice, as well as a dramatic increase in acetaminophen hepatotoxicity. In Cyp2e1(-/-) mice administered up to 600 mg acetaminophen/kg alone and euthanized 7 h later, there was no increase in serum levels of ALT. In Cyp2e1(-/-) mice pretreated with ethanol and isopentanol, subsequent exposure to 400 or 600 mg acetaminophen/kg resulted in centrilobular necrosis in all mice with maximal elevation in serum levels of ALT. Acetaminophen-mediated liver damage was similar in males and females. Hepatic microsomal levels of APAP activation in untreated females were similar to those in males treated with the alcohols. However, the females, like the males, required pretreatment with the alcohols in order to increase APAP hepatotoxicity. These findings suggest that, in the Cyp2e1(-/-) mice, the alcohol-mediated increase in acetaminophen hepatotoxicity involves the contribution of other factors, in addition to induction of CYP(s) that activate acetaminophen. Alternatively, CYP-mediated activation of acetaminophen measured in vitro may not reflect the actual activity in vivo. Our findings that a 7-day treatment with ethanol and isopentanol causes extensive hepatic steatosis and increases acetaminophen hepatotoxicity in Cyp2e(-/-) mice indicate that CYP2E1 is not essential for either response.

  7. The new ENSEMBLES E1 mitigation scenario for future climate simulations

    Science.gov (United States)

    Royer, J.-F.; Lowe, J.; Johns, T.; van Vuuren, D.; Stehfest, E.; Denoblet-Ducoudré, N.; Boucher, O.; Rognerud, B.; Huebener, H.

    2009-04-01

    that equivalent to 450-ppm of CO2 concentration. This new stabilisation scenario (E1) has been developed with the latest version of the integrated impact assessment model IMAGE at Netherlands Environmental Assessment Agency (MNP), with improvements in the land use representation of the A1B scenario. Though the coupled climate models are directly forced by the concentrations of the well-mixed greenhouse gases (CO2, CH4, N2O, CFCs and other minor species) produced by the IMAGE model, it has however been necessary to make further adaptations, by using additional modelling tools, in order to produce the other forcing data needed by the GCMs. The 3-dimensional atmospheric fields for anthropogenic sulphate aerosols concentrations have been computed by a chemistry-transport model using the emissions from the IMAGE scenarios. The ozone concentrations have been computed by the chemistry-transport model from the University of Oslo. The land use maps over the period 1850-2000 have been derived from the LUCID project based on a combination of the crop dataset of Ramankutty and Foley and pasture from the HYDE dataset. They have been then merged with the land-use maps produced by the IMAGE scenarios in order to derive adapted land-use maps over the 21st century. The different forcings and some results of the "stream 2" ENSEMBLES scenarios are described and illustrated in this presentation.

  8. CYP2A6 and CYP2E1 polymorphisms in a Brazilian population living in Rio de Janeiro

    Directory of Open Access Journals (Sweden)

    A. Rossini

    2006-02-01

    Full Text Available Cytochrome P450 (CYP is a superfamily of enzymes involved in the metabolism of endogenous compounds and xenobiotics. CYP2A6 catalyzes the oxidation of nicotine and the activation of carcinogens such as aflatoxin B1 and nitrosamines. CYP2E1 metabolizes ethanol and other low-molecular weight compounds and can also activate nitrosamines. The CYP2A6 and CYP2E1 genes are polymorphic, altering their catalytic activities and susceptibility to cancer and other diseases. A number of polymorphisms described are ethnic-dependent. In the present study, we determined the genotype and allele frequencies of the main CYP2A6 and CYP2E1 polymorphisms in a group of 289 volunteers recruited at the Central Laboratory of Hospital Universitário Pedro Ernesto. They had been residing in the city of Rio de Janeiro for at least 6 months and were divided into two groups according to skin color (white and non-white. The alleles were determined by allele specific PCR (CYP2A6 or by PCR-RFLP (CYP2E1. The frequencies of the CYP2A6*1B and CYP2A6*2 alleles were 0.29 and 0.02 for white individuals and 0.24 and 0.01 for non-white individuals, respectively. The CYP2A6*5 allele was not found in the population studied. Regarding the CYP2E1*5B allele, we found a frequency of 0.07 in white individuals, which was statistically different (P < 0.05 from that present in non-white individuals (0.03. CYP2E1*6 allele frequency was the same (0.08 in both groups. The frequencies of CYP2A6*1B, CYP2A6*2 and CYP2E1*6 alleles in Brazilians are similar to those found in Caucasians and African-Americans, but the frequency of the CYP2E1*5B allele is higher in Brazilians.

  9. The isolation of the ectodomain of the alphavirus E1 protein as a soluble hemagglutinin and its crystallization.

    Science.gov (United States)

    Wengler, G; Wengler, G; Rey, F A

    1999-05-10

    Alphaviruses are isometric enveloped viruses approximately 70 nm in diameter. The viral surface contains 80 glycoprotein spikes arranged in a T = 4 lattice. Each of these spikes consists of three heterodimers of the viral membrane proteins E1 (approximately 49 kDa) and E2 (approximately 51 kDa). Cryoelectron microscopic analyses have shown that the spikes form a protein shell on the viral surface. We have made an attempt to isolate biologically active protein fragments from this surface and to grow crystals from such fragments. To this end membrane proteins were extracted with Nonidet-P40 from the Semliki Forest alphavirus and the proteins were separated from detergent by centrifugation. A protein complex containing the E1 and E2 molecules in quantitative yield was obtained by this procedure. This complex has the following properties: It sediments at approximately 30S, it chromatographs with an apparent molecular mass of approximately 580,000 Da during gel filtration, it cannot be dissociated by either nonionic detergents or 6 M urea, and at acid pH it is a highly active hemagglutinin. The data indicate that this 30S hemagglutinin complex, which has not been hitherto described for alphaviruses, may represent a variant form of the protein lattice present on the alphavirus surface. Cleavage of this complex by subtilisin selectively removes carboxy-terminal sequences from the E1 and E2 proteins, which contain the cytoplasmic and transmembrane segments of the proteins and a small part of their ectodomain. The remaining ectodomains are called E1DeltaS and E2DeltaS. This proteolysis also leads to dissociation of the 30S complex. The cleavage products accumulate in the form of a heterodimer of the E1DeltaS and E2DeltaS proteins. Treatment of the heterodimer with PNGase F leads to rapid removal of carbohydrate from the E2DeltaS protein and a dissociation of the complex into the constituent molecules, which can be separated by chromatography. The finding that the

  10. Roles of CYP2e1 in 1,2-dichloroethane-induced liver damage in mice.

    Science.gov (United States)

    Sun, Qi; Wang, Gaoyang; Gao, Lanyue; Shi, Lei; Qi, Ying; Lv, Xiuqiang; Jin, Yaping

    2016-11-01

    The aim of this study was to explore the roles of cytochrome P450 2E1 (CYP2E1) in 1,2-dichloroethane (1,2-DCE)-induced liver damage. Two parts were included in this study: first, effect of 1,2-DCE on microsomal expression of CYP2E1, and second, potential of an inhibitor of CYP2E1 to reduce 1,2-DCE-induced liver damage. In part one, mice were exposed to 0, 0.225, 0.45, or 0.9 g/m(3) 1,2-DCE for 10 days, 3.5 h per day through static inhalation. In part two, mice were divided into blank control, solvent control, inhibitor control, 1,2-DCE-poisoned group, and low or high intervention group. In part one, compared to the control, serum alanine aminotransferase (ALT) activities and hepatic malondialdehyde (MDA) levels in 0.9 g/m(3) 1,2-DCE group, and microsomal CYP2E1 protein expression and activity in both 0.45 and 0.9 g/m(3) 1,2-DCE groups increased significantly; conversely, hepatic nonprotein sulfhydryl (NPSH) levels in both 0.45 and 0.9 g/m(3) 1,2-DCE groups and hepatic SOD activities in 0.9 g/m(3) 1,2-DCE group decreased significantly. In part two, microsomal CYP2E1 protein expression and activity decreased significantly in both low and high intervention groups compared to 1,2-DCE-poisoned group. Along with the changes of CYP2E1, hepatic MDA levels and serum ALT activities decreased; conversely, hepatic NPSH levels and SOD activities increased significantly in high intervention group. Taken together, our results suggested that 1,2-DCE could enhance CYP2E1 protein expression and enzymatic activity, which could cause oxidative damage in liver, serving as an important mechanism underlying 1,2-DCE-induced liver damage. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1430-1438, 2016.

  11. CYP2A6 and CYP2E1 polymorphisms in a Brazilian population living in Rio de Janeiro

    Directory of Open Access Journals (Sweden)

    Rossini A.

    2006-01-01

    Full Text Available Cytochrome P450 (CYP is a superfamily of enzymes involved in the metabolism of endogenous compounds and xenobiotics. CYP2A6 catalyzes the oxidation of nicotine and the activation of carcinogens such as aflatoxin B1 and nitrosamines. CYP2E1 metabolizes ethanol and other low-molecular weight compounds and can also activate nitrosamines. The CYP2A6 and CYP2E1 genes are polymorphic, altering their catalytic activities and susceptibility to cancer and other diseases. A number of polymorphisms described are ethnic-dependent. In the present study, we determined the genotype and allele frequencies of the main CYP2A6 and CYP2E1 polymorphisms in a group of 289 volunteers recruited at the Central Laboratory of Hospital Universitário Pedro Ernesto. They had been residing in the city of Rio de Janeiro for at least 6 months and were divided into two groups according to skin color (white and non-white. The alleles were determined by allele specific PCR (CYP2A6 or by PCR-RFLP (CYP2E1. The frequencies of the CYP2A6*1B and CYP2A6*2 alleles were 0.29 and 0.02 for white individuals and 0.24 and 0.01 for non-white individuals, respectively. The CYP2A6*5 allele was not found in the population studied. Regarding the CYP2E1*5B allele, we found a frequency of 0.07 in white individuals, which was statistically different (P < 0.05 from that present in non-white individuals (0.03. CYP2E1*6 allele frequency was the same (0.08 in both groups. The frequencies of CYP2A6*1B, CYP2A6*2 and CYP2E1*6 alleles in Brazilians are similar to those found in Caucasians and African-Americans, but the frequency of the CYP2E1*5B allele is higher in Brazilians.

  12. Ultrafast carrier relaxation through Auger recombination in the topological insulator B i1.5S b0.5T e1.7S e1.3

    Science.gov (United States)

    Onishi, Yoshito; Ren, Zhi; Segawa, Kouji; Kaszub, Wawrzyniec; Lorenc, Maciej; Ando, Yoichi; Tanaka, Koichiro

    2015-02-01

    Ultrafast carrier dynamics have great significance for our understanding of the transport properties of the surface state in topological insulator (TI) materials. We report midinfrared pump-probe measurements on the intrinsic TI material B i1.5S b0.5T e1.7S e1.3 and show that the change in photoinduced reflectivity can be decomposed into a fast negative part and a slow positive part. Calculations of the dielectric function made at various carrier temperatures and densities reveal that the fast negative component corresponds to the disappearance of the phase-space filling effect due to hot carriers around the probe energy and the decay component corresponds to the recombination of carriers near the band edge. The ratio of the fast negative component to the slow positive component is larger in the excitations conducted at the higher carrier densities, which suggests that the carrier temperature increases through Auger recombination. A qualitative analysis using rate equations reinforces this assumption, so we conclude that Auger recombination is the main cause of the population relaxation at carrier densities higher than 1018c m-3 and that we determined the Auger coefficient for B i1.5S b0.5T e1.7S e1.3 as C =0.4 ×10-26c m6/s .

  13. Iub接口部分E1中断对HSDPA吞吐率的影响及优化%Iub Interface Part E1 Interruption to HSDPA Throughput Rate Influence and Optimization

    Institute of Scientific and Technical Information of China (English)

    李筱林; 卢琦

    2013-01-01

    主述在3G网络中,Iub接口在基于ATM/E1的传输方式下,因为部分E1传输发生中断后,带宽分配无法动态调度,使实际网络资源能力小于规划配置资源需求从而引起瓶颈,导致数据传输过程中出现网络拥塞,使HSDPA产生吞吐率大幅度下降的情况。通过剖析其内在拥塞原因,研究其拥塞控制优化手段,从而提高用户的感受度。%Mainly describes the 3G network, Iub interface based ATM/E1 transmission, because part of the E1 transmis-sion interruption, bandwidth allocation can not be dynamic scheduling, planning capacity less than the actual network re-sources causing the bottleneck resource allocation needs, resulting inthe digital transmission network congestion, the HS-DPA throughput dropped significantly; By analyzing its inherent congestion reasons to study its congestion control opti-mization tools to improve the user's feelings.

  14. Crystal structure of (E-1-[4-({4-[(4-methoxybenzylideneamino]phenyl}sulfanylphenyl]ethan-1-one

    Directory of Open Access Journals (Sweden)

    Rabihe Hebbachi

    2015-02-01

    Full Text Available The title Schiff base compound, C22H19NO2S, crystallized with two independent molecules (A and B in the asymmetric unit. Both molecules have an E conformation about the C=N bond. The two molecules differ in the orientation of the aromatic rings with respect to each other. The outer 4-methoxybenzene ring is inclined to the central benzene ring and the outer 4-acetylbenzene ring by 1.80 (19 and 63.73 (19°, respectively, in molecule A, and by 6.72 (18 and 68.53 (19°, respectively, in molecule B. The two outer benzene rings are inclined to one another by 63.77 (18 and 63.19 (18° in molecules A and B, respectively. In the crystal, the individual molecules stack in columns along [010], and are linked by a number of C—H...π interactions, forming slabs lying parallel to (001.

  15. [Conservation Units.

    Science.gov (United States)

    Texas Education Agency, Austin.

    Each of the six instructional units deals with one aspect of conservation: forests, water, rangeland, minerals (petroleum), and soil. The area of the elementary school curriculum with which each correlates is indicated. Lists of general and specific objectives are followed by suggested teaching procedures, including ideas for introducing the…

  16. [Conservation Units.

    Science.gov (United States)

    Texas Education Agency, Austin.

    Instructional units deal with each aspect of conservation: forests, wildlife, rangelands, water, minerals, and soil. The area of the secondary school curriculum with which each is correlated is indicated. Lists of general and specific objectives are followed by suggested teaching procedures, including ideas for introducing the topic, questions to…

  17. Preclinical safety assessment of Ad[I/PPT-E1A], a novel oncolytic adenovirus for prostate cancer.

    Science.gov (United States)

    Schenk, Ellen; Essand, Magnus; Kraaij, Robert; Adamson, Rachel; Maitland, Norman J; Bangma, Chris H

    2014-03-01

    Prostate cancer is the most common malignancy in the Western world. Patients can be cured only when the tumor has not metastasized outside the prostate. However, treatment with curative intent fails in a significant number of men, often resulting in untreatable progressive disease with a fatal outcome. Oncolytic adenovirus therapy may be a promising adjuvant treatment to reduce local failure or the outgrowth of micrometastatic disease. Within the European gene therapy consortium GIANT, we have developed a novel prostate-specific oncolytic adenovirus: Ad[I/PPT-E1A]. This adenovirus specifically kills prostate cells via prostate-specific replication. This article describes the clinical development of Ad[I/PPT-E1A] with particular reference to the preclinical safety assessment of this novel virus. The preclinical safety assessment involved an efficacy study in a human orthotopic xenograft mouse model, a specificity study in human primary cells, and a toxicity study in normal mice. These studies confirmed that Ad[I/PPT-E1A] efficiently kills prostate tumor cells in vivo, is not harmful to other organs, and is well tolerated in mice after systemic delivery. The safety, as well as the immunological effects of Ad[I/PPT-E1A] as a local adjuvant therapy, will now be studied in a phase I dose-escalating trial in patients with localized prostate cancer who are scheduled for curative radical prostatectomy and can be used as an updated paradigm for similar therapeutic viruses.

  18. Pre-clinical safety assessment of Ad[I/PPT-E1A], a novel oncolytic adenovirus for prostate cancer.

    Science.gov (United States)

    Schenk, Ellen; Essand, Magnus; Kraaij, Robert; Adamson, Rachel; Maitland, Norman; Bangma, Chris H

    2014-02-18

    Prostate cancer is the most common malignancy in the Western world. Patients can only be cured when the tumour has not metastasised outside the prostate. However, treatment with curative intent fails in a significant number of men, often resulting in untreatable progressive disease with a fatal outcome. Oncolytic adenovirus therapy may be a promising adjuvant treatment to reduce local failure or the outgrowth of micrometastatic disease. Within the European gene therapy consortium GIANT, we have developed a novel prostate-specific oncolytic adenovirus: Ad[I/PPT-E1A] that specifically kills prostate cells via prostate-specific replication. This paper describes the clinical development of Ad[I/PPT-E1A] with particular reference to the pre-clinical safety assessment of this novel virus. The pre-clinical safety assessment involved an efficacy study in a human orthotopic xenograft mouse model, a specificity study in human primary cells and a toxicity study in normal mice. These studies confirmed that Ad[I/PPT-E1A] efficiently kills prostate tumour cells in vivo, is not harmful to other organs and is well-tolerated in mice after systemic delivery. The safety, as well as the immunological effects of Ad[I/PPT-E1A] as a local adjuvant therapy will now be studied in a phase I dose-escalating trial in patients with localised prostate cancer who are scheduled for curative radical prostatectomy and can be used as an updated paradigm for similar therapeutic viruses.

  19. Preface: Joint researches are benefiting the Chang'E-1 comprehensive lunar scientific studies which probe ever deeper

    Institute of Scientific and Technical Information of China (English)

    PING JinSong

    2010-01-01

    @@ Payloads of the first Chinese lunar mission Chang'E-1 obtained fruitful scientific data which cover a wide range of disci plines and fields.From 2007, many domestic research teams have been working hard on calibrating and validating these data cautiously and carefully.

  20. Identification and Characteristics of a Novel E1 Like Gene nUBE1L in Human Testis

    Institute of Scientific and Technical Information of China (English)

    Hui ZHU; Jia-Hao SHA; Zuo-Min ZHOU; Ran HUO; Xiao-Yan HUANG; Li LU; Min LIN; Li-Rong WANG; Ya-Dong ZHOU; Jian-Min LI

    2004-01-01

    A gene,presumably involved in spermatogenesis,was identified and characterized by using cDNA microarray.Hybridization intensity was 2.13 fold higher in adult testis than that in fetal testis.The full length of this gene was 4288 bp and it encoded a 578 amino acid protein.Conserved structure and amino acid sequence analysis revealed that the protein contained 1 Thif-domain,2 UBACT-domains,and a functional active site cysteine lay upstream of UBACT domain,all of them also existed in ubiquitinactivating enzyme E1 and E1 like proteins.So we named this gene as a novel ubiquitin-activating enzyme E1 like gene(nUBE1L).Expression profile showed that nUBEIL was predominantly expressed in testis.Comparison of the expression of nUBE1L in different developmental stages of testis indicated that it was highly expressed in adult testis.In conclusion,nUBE1L is a novel human E1 like gene highly expressed in adult testis,which plays key role in ubiquitin system,and accordingly influences spermatogenesis and male fertility.

  1. Structural basis of steroid binding and oxidation by the cytochrome P450 CYP109E1 from Bacillus megaterium

    NARCIS (Netherlands)

    Jozwik, Ilona; Kiss, Flora M; Gricman, Łukasz; Abdulmughni, Ammar; Brill, Elisa; Zapp, Josef; Pleiss, Juergen; Bernhardt, Rita; Thunnissen, Andy-Mark W H

    2016-01-01

    Cytochrome P450 monooxygenases (P450s) are attractive enzymes for the pharmaceutical industry, in particular for applications in steroidal drug synthesis. Here we report a comprehensive functional and structural characterization of CYP109E1, a novel steroid-converting cytochrome P450 enzyme identifi

  2. Salmonid alphavirus glycoprotein E2 requires low temperature and E1 for virion formation and induction of protective immunity.

    Science.gov (United States)

    Hikke, Mia C; Braaen, Stine; Villoing, Stephane; Hodneland, Kjartan; Geertsema, Corinne; Verhagen, Lisa; Frost, Petter; Vlak, Just M; Rimstad, Espen; Pijlman, Gorben P

    2014-10-29

    Salmonid alphavirus (SAV; also known as Salmon pancreas disease virus; family Togaviridae) causes pancreas disease and sleeping disease in Atlantic salmon and rainbow trout, respectively, and poses a major burden to the aquaculture industry. SAV infection in vivo is temperature-restricted and progeny virus is only produced at low temperatures (10-15 °C). Using engineered SAV replicons we show that viral RNA replication is not temperature-restricted suggesting that the viral structural proteins determine low-temperature dependency. The processing/trafficking of SAV glycoproteins E1 and E2 as a function of temperature was investigated via baculovirus vectors in Sf9 insect cells and by transfection of CHSE-214 fish cells with DNA constructs expressing E1 and E2. We identified SAV E2 as the temperature determinant by demonstrating that membrane trafficking and surface expression of E2 occurs only at low temperature and only in the presence of E1. Finally, a vaccination-challenge model in Atlantic salmon demonstrates the biological significance of our findings and shows that SAV replicon DNA vaccines encoding E2 elicit protective immunity only when E1 is co-expressed. This is the first study that identifies E2 as the critical determinant of SAV low-temperature dependent virion formation and defines the prerequisites for induction of a potent immune response in Atlantic salmon by DNA vaccination.

  3. Prostaglandin E1 treatment in ductus dependent congenital cardiac malformation. A review of the treatment of 34 neonates

    DEFF Research Database (Denmark)

    Høst, A; Halken, S; Kamper, J

    1988-01-01

    Thirty-four sick neonates with major duct dependent cardiac defects were given short term (1 h-408 h) intravenous infusions of prostaglandin E1 (alprostadil) in doses varying between 0.1 micrograms/kg/min (starting dose) and 0.01 micrograms/kg/min. The aim of the study was to establish an effecti...

  4. How the MccB bacterial ancestor of ubiquitin E1 initiates biosynthesis of the microcin C7 antibiotic

    Energy Technology Data Exchange (ETDEWEB)

    Regni, Catherine A.; Roush, Rebecca F.; Miller, Darcie J.; Nourse, Amanda; Walsh, Christopher T.; Schulman, Brenda A.; (Harvard-Med); (HHMI); (SJCH)

    2009-09-11

    The 39-kDa Escherichia coli enzyme MccB catalyses a remarkable posttranslational modification of the MccA heptapeptide during the biosynthesis of microcin C7 (MccC7), a 'Trojan horse' antibiotic. The approximately 260-residue C-terminal region of MccB is homologous to ubiquitin-like protein (UBL) activating enzyme (E1) adenylation domains. Accordingly, MccB-catalysed C-terminal MccA-acyl-adenylation is reminiscent of the E1-catalysed activation reaction. However, unlike E1 substrates, which are UBLs with a C-terminal di-glycine sequence, MccB's substrate, MccA, is a short peptide with an essential C-terminal Asn. Furthermore, after an intramolecular rearrangement of MccA-acyl-adenylate, MccB catalyses a second, unique reaction, producing a stable phosphoramidate-linked analogue of acyl-adenylated aspartic acid. We report six-crystal structures of MccB in apo, substrate-, intermediate-, and inhibitor-bound forms. Structural and kinetic analyses reveal a novel-peptide clamping mechanism for MccB binding to heptapeptide substrates and a dynamic-active site for catalysing dual adenosine triphosphate-consuming reactions. The results provide insight into how a distinctive member of the E1 superfamily carries out two-step activation for generating the peptidyl-antibiotic MccC7.

  5. Mobilization properties of small ColE1-like plasmids carrying kanamycin resistance gene isolated from Salmonella enterica serotypes

    Science.gov (United States)

    Background: Previously we isolated and characterized various groups of small kanamycin resistance (KanR) ColE1-like plasmids from different serotypes of Salmonella enterica isolates. These plasmids all carried the aph(3)-I gene encoding the aminoglycoside phosphotransferase responsible for the kanam...

  6. 26 CFR 301.6503(e)-1 - Suspension of running of period of limitation; certain powers of appointment.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 18 2010-04-01 2010-04-01 false Suspension of running of period of limitation... Limitations Limitations on Assessment and Collection § 301.6503(e)-1 Suspension of running of period of... complied with, the running of the period of limitation for assessment or collection of any estate...

  7. The making of S{u00E1mi ethnography : contested authorities and negotiated representations / Kristin Kuutma

    Index Scriptorium Estoniae

    Kuutma, Kristin, 1959-

    2008-01-01

    Vaadeldakse saami autori Johan Turi raamatut Muitalus s{u00E1miid birra (1910) ja selle tõlget taani keelede Emilie Demant Hatti poolt ning uuritakse, millist autoritaarsust ja kultuurilist poeetikat see peegeldab ja millist sotsiaalset ja kultuurilist kriitikat ta edasi arendab

  8. The making of S{u00E1mi ethnography : contested authorities and negotiated representations / Kristin Kuutma

    Index Scriptorium Estoniae

    Kuutma, Kristin, 1959-

    2008-01-01

    Vaadeldakse saami autori Johan Turi raamatut Muitalus s{u00E1miid birra (1910) ja selle tõlget taani keelede Emilie Demant Hatti poolt ning uuritakse, millist autoritaarsust ja kultuurilist poeetikat see peegeldab ja millist sotsiaalset ja kultuurilist kriitikat ta edasi arendab

  9. The pharmacological effects of morroniside and loganin isolated from Liuweidihuang Wan, on MC3T3-E1 cells.

    Science.gov (United States)

    Li, Manyu; Wang, Wei; Wang, Ping; Yang, Kun; Sun, Hui; Wang, Xijun

    2010-10-21

    Liuweidihuang wan (LW), initially a well-known formula for curing "wu chi wu ruan", is commonly used nowadays for clinical treatment of postmenopausal osteoporosis (PO), but the identity of the effective substance(s) remains unclear. The present study was designed to evaluate the effects of morroniside and loganin isolated from LW on the proliferation, differentiation and apoptosis of MC3T3-E1 cells, as well as the possible mechanism of action. Morroniside and loganin had no effects on the proliferation of MC3T3-E1 cells, but both susbtances could improve the activity of alkaline phosphatase (ALP), and increase the contents of collagen type I and osteocalcin. Simultaneously, the mRNA expression of caspase-3, capase-9, RANKL was down-regulated and that of bcl-2 was up-regulated, which partially explains the anti-osteoporosis mechanism in MC3T3-E1 cells. In conclusion, morroniside and loganin may directly promote the differentiation and inhibit the apoptosis of MC3T3-E1 cells, and accordingly indirectly reduce bone resorption, which makes them promising natural drugs leads for treating PO in the near future.

  10. The Pharmacological Effects of Morroniside and Loganin Isolated from Liuweidihuang Wan, on MC3T3-E1 Cells

    Directory of Open Access Journals (Sweden)

    Xijun Wang

    2010-10-01

    Full Text Available Liuweidihuang Wan (LW, initially a well-known formula for curing “wu chi wu ruan”, is commonly used nowadays for clinical treatment of Postmenopausal Osteoporosis (PO, but the identity of the effective substance(s remains unclear. The present study was designed to evaluate the effects of morroniside and loganin isolated from LW on the proliferation, differentiation and apoptosis of MC3T3-E1 cells, as well as the possible mechanism of action. Morroniside and loganin had no effects on the proliferation of MC3T3-E1 cells, but both susbtances could improve the activity of alkaline phosphatase (ALP, and increase the contents of collagen type I and osteocalcin. Simultaneously, the mRNA expression of caspase-3, capase-9, RANKL was down-regulated and that of bcl-2 was up-regulated, which partially explains the anti-osteoporosis mechanism in MC3T3-E1 cells. In conclusion, morroniside and loganin may directly promote the differentiation and inhibit the apoptosis of MC3T3-E1 cells, and accordingly indirectly reduce bone resorption, which makes them promising natural drugs leads for treating PO in the near future.

  11. Comparison of T7E1 and surveyor mismatch cleavage assays to detect mutations triggered by engineered nucleases.

    Science.gov (United States)

    Vouillot, Léna; Thélie, Aurore; Pollet, Nicolas

    2015-01-07

    Genome editing using engineered nucleases is used for targeted mutagenesis. But because genome editing does not target all loci with similar efficiencies, the mutation hit-rate at a given locus needs to be evaluated. The analysis of mutants obtained using engineered nucleases requires specific methods for mutation detection, and the enzyme mismatch cleavage method is used commonly for this purpose. This method uses enzymes that cleave heteroduplex DNA at mismatches and extrahelical loops formed by single or multiple nucleotides. Bacteriophage resolvases and single-stranded nucleases are used commonly in the assay but have not been compared side-by-side on mutations obtained by engineered nucleases. We present the first comparison of the sensitivity of T7E1 and Surveyor EMC assays on deletions and point mutations obtained by zinc finger nuclease targeting in frog embryos. We report the mutation detection limits and efficiencies of T7E1 and Surveyor. In addition, we find that T7E1 outperforms the Surveyor nuclease in terms of sensitivity with deletion substrates, whereas Surveyor is better for detecting single nucleotide changes. We conclude that T7E1 is the preferred enzyme to scan mutations triggered by engineered nucleases.

  12. Intra-arterial infusion of prostaglandin E1 in normal subjects and patients with peripheral arterial disease

    DEFF Research Database (Denmark)

    Nielsen, P E; Nielsen, S L; Holstein, P;

    1976-01-01

    Acute vasodilatation was produced by infusion of prostaglandin E1 (PGE1) in the femoral artery in 6 patients with occlusive arterial disease of the legs and in 3 normal subjects. The effect on blood flow and on blood pressure was measured at different segments of the leg with the strain gauge...

  13. Lokal østrogen-fobehandling reducerer aborttiden ven prostaglandin E1 analog-induceret abort i 2. trimester

    DEFF Research Database (Denmark)

    Petersen, Lone Kjeld; Uldbjerg, N; Allen, J G

    1991-01-01

    Twenty-eight patients (median gestational age 17 weeks) referred for induction of second trimester abortion, were randomized to intracervical preliminary treatment by either 50 mg 17 beta-oestradiol or placebo. Abortion was then induced by 1 mg prostaglandin E1 vagitories. The preliminary treatme...

  14. Structural and Dynamic Characterization of the Molecular Hub Early Region 1A (E1A) from Human Adenovirus.

    Science.gov (United States)

    Hošek, Tomáš; Calçada, Eduardo O; Nogueira, Marcela Oliveira; Salvi, Michele; Pagani, Talita Duarte; Felli, Isabella C; Pierattelli, Roberta

    2016-09-05

    The small-DNA human adenovirus encodes one of the most versatile molecular hubs, the E1A protein. This protein is essential for productive viral infection in human cells and a vast amount of biologically relevant data are available on its interactions with host proteins. Up to now, however, no high-resolution structural and dynamic information on E1A is available despite its important biological role. Among the different spliced variants of E1A, two are expressed at high level in the early stage of infection. These are 243 and 289 residues isoforms. Herein, we present their NMR characterization, showing that they are both highly disordered, but also demonstrate a certain heterogeneous behavior in terms of structural and dynamic properties. Furthermore, we present the characterization of the isolated domain of the longer variant, known as CR3. This study opens the way to understanding at the molecular level how E1A functions. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. CYP2E1 Rsa Ⅰ polymorphism impacts on risk of colorectal cancer association with smoking and alcohol drinking

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To investigate associations between the Rsa Ⅰpolymorphism of CYP2E1 and risk of colorectal cancer.METHODS: A case-control study was conducted with 315 colorectal cancer cases (105 colon, 210 rectal)and 439 population-based controls in Jiangsu Province of China. Genomic DNA samples were assayed for restriction fragment length polymorphisms in CYP2E1by PCR amplification followed by digestion with Rsa Ⅰ. Information on smoking and alcohol drinking was collected using a questionnaire. Odds ratios (ORs) were estimated with an unconditional logistic model.RESULTS: The proportional distribution of the CYP2E1 Rsa Ⅰ c1/c1, c1/c2 and c2/c2 genotypes were 61.4%,35.6% and 3.0% in controls, 60.6%, 33.7% and 5.8%in colon cancer cases, and 58.4%, 34.0% and 7.7% in rectal cancer cases, respectively. A significant difference was noted between controls and rectal cancer cases (P = 0.029), the c2/c2 genotype being associated with elevated OR (adjusted age, sex and status of the smoking and alcohol drinking) for rectal cancer (1.64,95% CI, 1.12-2.41, vs c1 allele carriers), but not for colon cancer. In interaction analysis between the CYP2E1Rsa Ⅰ genotype and smoking and drinking habits, we found a significant cooperative action between the c2/c2 genotype and alcohol drinking in the sex-, age-adjusted ORs for both colon (4.74, 95% CI, 1.10-20.40) and rectal (5.75, 95% CI, 1.65-20.05) cancers. Among nonsmokers, the CYP2E1 Rsa Ⅰ c2/c2 genotype was also associated with elevated ORs in the two sites (1.95, 95%CI, 0.99-3.86 and 2.30, 95% CI, 1.32-3.99).CONCLUSION: The results of the present study suggest that the CYP2E1 c2/c2 genotype increases susceptibility to rectal cancer and the gene-environmental interactions between the CYP2E1 polymorphism and smoking or alcohol drinking exist for colorectal neoplasia in general.

  16. Ghrelin inhibits the apoptosis of MC3T3-E1 cells through ERK and AKT signaling pathway

    Energy Technology Data Exchange (ETDEWEB)

    Liang, Qiu-Hua; Liu, Yuan; Wu, Shan-Shan; Cui, Rong-Rong; Yuan, Ling-Qing, E-mail: allenylq@hotmail.com; Liao, Er-Yuan, E-mail: eyliao@21cn.com

    2013-11-01

    Ghrelin is a 28-amino-acid peptide that acts as a natural endogenous ligand of the growth hormone secretagogue receptor (GHSR) and strongly stimulates the release of growth hormone from the hypothalamus–pituitary axis. Previous studies have identified the important physiological effects of ghrelin on bone metabolism, such as regulating proliferation and differentiation of osteoblasts, independent of GH/IGF-1 axis. However, research on effects and mechanisms of ghrelin on osteoblast apoptosis is still rare. In this study, we identified expression of GHSR in MC3T3-E1 cells and determined the effects of ghrelin on the apoptosis of osteoblastic MC3T3-E1 cells and the mechanism involved. Our data demonstrated that ghrelin inhibited the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, as determined by terminal deoxynucleotidyl transferase-mediated deoxyribonucleotide triphosphate nick end-labeling (TUNEL) and ELISA assays. Moreover, ghrelin upregulated Bcl-2 expression and downregulated Bax expression in a dose-dependent manner. Our study also showed decreased activated caspase-3 activity under the treatment of ghrelin. Further study suggested that ghrelin stimulated the phosphorylation of ERK and AKT. Pretreatment of cells with the ERK inhibitor PD98059, PI3K inhibitor LY294002, and GHSR-siRNA blocked the ghrelin-induced activation of ERK and AKT, respectively; however, ghrelin did not stimulate the phosphorylation of p38 or JNK. PD90859, LY294002 and GHSR-siRNA attenuated the anti-apoptosis effect of ghrelin in MC3T3-E1 cells. In conclusion, ghrelin inhibits the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, which may be mediated by activating the GHSR/ERK and GHSR/PI3K/AKT signaling pathways. - Highlights: • We explored the effects of ghrelin on serum deprivation-induced MC3T3-E1 cells apoptosis. • Both ELISA and TUNEL were used to detect the apoptosis. • The receptor of ghrelin, GHSR, was expressed in MC3T3-E1

  17. Monothiol glutaredoxin-BolA interactions: redox control of Arabidopsis thaliana BolA2 and SufE1.

    Science.gov (United States)

    Couturier, Jérémy; Wu, Hui-Chen; Dhalleine, Tiphaine; Pégeot, Henri; Sudre, Damien; Gualberto, José M; Jacquot, Jean-Pierre; Gaymard, Frédéric; Vignols, Florence; Rouhier, Nicolas

    2014-01-01

    A functional relationship between monothiol glutaredoxins and BolAs has been unraveled by genomic analyses and in several high-throughput studies. Phylogenetic analyses coupled to transient expression of green fluorescent protein (GFP) fusions indicated that, in addition to the sulfurtransferase SufE1, which contains a C-terminal BolA domain, three BolA isoforms exist in Arabidopsis thaliana, BolA1 being plastidial, BolA2 nucleo-cytoplasmic, and BolA4 dual-targeted to mitochondria and plastids. Binary yeast two-hybrid experiments demonstrated that all BolAs and SufE1, via its BolA domain, can interact with all monothiol glutaredoxins. Most interactions between protein couples of the same subcellular compartment have been confirmed by bimolecular fluorescence complementation. In vitro experiments indicated that monothiol glutaredoxins could regulate the redox state of BolA2 and SufE1, both proteins possessing a single conserved reactive cysteine. Indeed, a glutathionylated form of SufE1 lost its capacity to activate the cysteine desulfurase, Nfs2, but it is reactivated by plastidial glutaredoxins. Besides, a monomeric glutathionylated form and a dimeric disulfide-bridged form of BolA2 can be preferentially reduced by the nucleo-cytoplasmic GrxS17. These results indicate that the glutaredoxin-BolA interaction occurs in several subcellular compartments and suggest that a redox regulation mechanism, disconnected from their capacity to form iron-sulfur cluster-bridged heterodimers, may be physiologically relevant for BolA2 and SufE1.

  18. Structural Basis for Cooperative Binding of Azoles to CYP2E1 as Interpreted through Guided Molecular Dynamics Simulations

    Science.gov (United States)

    Levy, Joseph W.; Hartman, Jessica H.; Perry, Martin D.; Miller, Grover P.

    2015-01-01

    CYP2E1 metabolizes a wide array of small, hydrophobic molecules, resulting in their detoxification or activation into carcinogens through Michaelis-Menten as well as cooperative mechanisms. Nevertheless, the molecular determinants for CYP2E1 specificity and metabolic efficiency toward these compounds are still unknown. Herein, we employed computational docking studies coupled to Molecular Dynamics simulations to provide a critical perspective for understanding a structural basis for cooperativity observed for an array of azoles from our previous binding and catalytic studies (Hartman, JH et al (2014) Biochem Pharmacol 87, 523-33). The resulting 28 CYP2E1 complexes in this study revealed a common passageway for azoles that included a hydrophobic steric barrier causing a pause in movement toward the active site. The entrance to the active site acted like a second sieve to restrict access to the inner chamber. Collectively, these interactions impacted the final orientation of azoles reaching the active site and hence could explain differences in their biochemical properties observed in our previous studies, such as the consequences of methylation at position 5 of the azole ring. The association of a second azole demonstrated significant differences in interactions stabilizing the bound complex than observed for the first binding event. Intermolecular interactions occurred between the two azoles as well as CYP2E1 residue side chains and backbone and involved both hydrophobic contacts and hydrogen bonds. The relative importance of these interactions depended on the structure of the respective azoles indicating the absence of specific defining criteria for binding unlike the well-characterized dominant role of hydrophobicity in active site binding. Consequently, the structure activity relationships described here and elsewhere are necessary to more accurately identify factors impacting the observation and significance of cooperativity in CYP2E1 binding and catalysis

  19. Role of CYP2E1-mediated metabolism in the acute and vestibular toxicities of nineteen nitriles in the mouse.

    Science.gov (United States)

    Saldaña-Ruíz, Sandra; Soler-Martín, Carla; Llorens, Jordi

    2012-01-25

    Allylnitrile, cis-crotononitrile, and 3,3'-iminodipropionitrile are known to cause vestibular toxicity in rodents, and evidence is available indicating that cis-2-pentenenitrile shares this effect. We evaluated nineteen nitriles for vestibular toxicity in wild type (129S1) and CYP2E1-null mice, including all the above, several neurotoxic nitriles, and structurally similar nitriles. A new acute toxicity test protocol was developed to facilitate evaluation of the vestibular toxicity by a specific behavioral test battery at doses up to sub-lethal levels while using a limited number of animals. A mean number of 8.5±0.3 animals per nitrile, strain and sex was necessary to obtain evidence of vestibular toxicity and optionally an estimation of the lethal dose. For several but not all nitriles, lethal doses significantly increased in CYP2E1-null mice. The protocol revealed the vestibular toxicity of five nitriles, including previously identified ototoxic compounds and one nitrile (trans-crotononitrile) known to have a different profile of neurotoxic effects in the rat. In all five cases, both sexes were affected and no decrease in susceptibility was apparent in CYP2E1-null mice respect to 129S1 mice. Fourteen nitriles caused no vestibular toxicity, including six nitriles tested in CYP2E1-null mice at doses significantly larger than the maximal doses that can be tested in wild type animals. We conclude that only a subset of low molecular weight nitriles is toxic to the vestibular system, that species-dependent differences exist in this vestibular toxicity, and that CYP2E1-mediated metabolism is not involved in this effect of nitriles although it has a role in the acute lethality of some of these compounds. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  20. A negative cooperativity mechanism of human CYP2E1 inferred from molecular dynamics simulations and free energy calculations.

    Science.gov (United States)

    Li, Jue; Wei, Dong-Qing; Wang, Jing-Fang; Li, Yi-Xue

    2011-12-27

    Human cytochrome P450 2E1 (CYP2E1) participates in the metabolism of over 2% of all the oral drugs. A hallmark peculiar feature of this enzyme is that it exhibits a pronounced negative cooperativity in substrate binding. However the mechanism by which the negative cooperativity occurs is unclear. Here, we performed molecular dynamics simulations and free energy calculations on human CYP2E1 to examine the structural differences between the substrate-free and the enzymes with one and two aniline molecules bound. Our results indicate that although the effector substrate does not bind in the active site cavity, it still can directly interact with the active site residues of human CYP2E1. The interaction of the effector substrate with the active site leads to a reorientation of active site residues, which thereby weakens the interactions of the active substrate with this site. We also identify a conserved residue T303 that plays a crucial role in the negative cooperative binding on the short-range effects. This residue is a key factor in the positioning of substrates and in proton delivery to the active site. Additionally, a long-range effect of the effector substrate is identified in which F478 is proposed to play a key role. As located in the interface between the active and effector sites, this residue structurally links the active and effector sites and is found to play a significant role in affecting substrate access and ligand positioning within the active site. In the negative cooperative binding, this residue can decrease the interactions of the active substrate with the active site by π-π stacking which then lowers the hydroxylation activity for the active substrate. These findings are in agreement with previous experimental observations and thus provide detailed atomistic insight into the poorly understood mechanism of the negative cooperativity in human CYP2E1.

  1. Crystal Structure of UBA2[superscript ufd]-Ubc9: Insights into E1-E2 Interactions in Sumo Pathways

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jing; Taherbhoy, Asad M.; Hunt, Harold W.; Seyedin, Steven N.; Miller, David W.; Miller, Darcie J.; Huang, Danny T.; Schulman, Brenda A. (SJCH)

    2012-04-30

    Canonical ubiquitin-like proteins (UBLs) such as ubiquitin, Sumo, NEDD8, and ISG15 are ligated to targets by E1-E2-E3 multienzyme cascades. The Sumo cascade, conserved among all eukaryotes, regulates numerous biological processes including protein localization, transcription, DNA replication, and mitosis. Sumo conjugation is initiated by the heterodimeric Aos1-Uba2 E1 enzyme (in humans called Sae1-Uba2), which activates Sumo's C-terminus, binds the dedicated E2 enzyme Ubc9, and promotes Sumo C-terminal transfer between the Uba2 and Ubc9 catalytic cysteines. To gain insights into details of E1-E2 interactions in the Sumo pathway, we determined crystal structures of the C-terminal ubiquitin fold domain (ufd) from yeast Uba2 (Uba2{sup ufd}), alone and in complex with Ubc9. The overall structures of both yeast Uba2{sup ufd} and Ubc9 superimpose well on their individual human counterparts, suggesting conservation of fundamental features of Sumo conjugation. Docking the Uba2{sup ufd}-Ubc9 and prior full-length human Uba2 structures allows generation of models for steps in Sumo transfer from Uba2 to Ubc9, and supports the notion that Uba2 undergoes remarkable conformational changes during the reaction. Comparisons to previous structures from the NEDD8 cascade demonstrate that UBL cascades generally utilize some parallel E1-E2 interaction surfaces. In addition, the structure of the Uba2{sup ufd}-Ubc9 complex reveals interactions unique to Sumo E1 and E2. Comparison with a previous Ubc9-E3 complex structure demonstrates overlap between Uba2 and E3 binding sites on Ubc9, indicating that loading with Sumo and E3-catalyzed transfer to substrates are strictly separate steps. The results suggest mechanisms establishing specificity and order in Sumo conjugation cascades.

  2. Sulforaphane Induces Nrf2 and Protects Against CYP2E1-dependent Binge Alcohol –induced Liver Steatosis

    Science.gov (United States)

    Zhou, Richard; Lin, Jianjun; Wu, Defeng

    2013-01-01

    Background The mechanism(s) by which alcohol causes cell injury are still not clear but a major mechanism appears to be the role of lipid peroxidation and oxidative stress in alcohol toxicity. CYP2E1-generated ROS contributes to the ethanol-induced oxidant stress and inhibition of CYP2E1 activity decreases ethanol-induced fatty liver. The transcription factor Nrf2 regulates the expression of many cytoprotective enzymes which results in cellular protection against a variety of toxins. Method The current study was designed to evaluate the ability of sulforaphane, an activator of Nrf2, to blunt CYP2E1-dependent, ethanol-induced steatosis in vivo and in vitro. Results The sulforaphane treatment activated Nrf2, increased levels of the Nrf2 target heme oxygenase -1 and subsequently lowered oxidant stress as shown by the decline in lipid peroxidation and 3-Nitrotyrosine protein adducts and an increase in GSH levels after the acute ethanol treatment. It decreased ethanol-elevated liver levels of triglycerides and cholesterol and Oil Red O staining. Similar results were found in vitro as addition of sulforaphane to HepG2 E47 cells, which express CYP2E1, elevated Nrf2 levels and decreased the accumulation of lipid in cells cultured with ethanol. Sulforaphane treatment had no effect on levels of or activity of CYP2E1. Conclusions Sulforaphane proved to be an effective in vivo inhibitor of acute ethanol–induced fatty liver in mice. General significance The possible amelioration of liver injury which occurs under these conditions by chemical activators of Nrf2 is of clinical relevance and worthy of further study. PMID:24060752

  3. Sulforaphane induces Nrf2 and protects against CYP2E1-dependent binge alcohol-induced liver steatosis.

    Science.gov (United States)

    Zhou, Richard; Lin, Jianjun; Wu, Defeng

    2014-01-01

    The mechanism(s) by which alcohol causes cell injury are still not clear but a major mechanism appears to be the role of lipid peroxidation and oxidative stress in alcohol toxicity. CYP2E1-generated ROS contributes to the ethanol-induced oxidant stress and inhibition of CYP2E1 activity decreases ethanol-induced fatty liver. The transcription factor Nrf2 regulates the expression of many cytoprotective enzymes which results in cellular protection against a variety of toxins. The current study was designed to evaluate the ability of sulforaphane, an activator of Nrf2, to blunt CYP2E1-dependent, ethanol-induced steatosis in vivo and in vitro. The sulforaphane treatment activated Nrf2, increased levels of the Nrf2 target heme oxygenase-1 and subsequently lowered oxidant stress as shown by the decline in lipid peroxidation and 3-nitrotyrosine protein adducts and an increase in GSH levels after the acute ethanol treatment. It decreased ethanol-elevated liver levels of triglycerides and cholesterol and Oil Red O staining. Similar results were found in vitro as addition of sulforaphane to HepG2 E47 cells, which express CYP2E1, elevated Nrf2 levels and decreased the accumulation of lipid in cells cultured with ethanol. Sulforaphane treatment had no effect on levels of or activity of CYP2E1. Sulforaphane proved to be an effective in vivo inhibitor of acute ethanol-induced fatty liver in mice. The possible amelioration of liver injury which occurs under these conditions by chemical activators of Nrf2 is of clinical relevance and worthy of further study. © 2013.

  4. 26 CFR 1.417(e)-1T - Restrictions and valuations of distributions from plans subject to sections 401(a)(11) and 417...

    Science.gov (United States)

    2010-04-01

    ... from plans subject to sections 401(a)(11) and 417. (Temporary) 1.417(e)-1T Section 1.417(e)-1T Internal... TAXES Pension, Profit-Sharing, Stock Bonus Plans, Etc. § 1.417(e)-1T Restrictions and valuations of distributions from plans subject to sections 401(a)(11) and 417. (Temporary) (a) (b) Consent, etc....

  5. 26 CFR 1.860E-1 - Treatment of taxable income of a residual interest holder in excess of daily accruals.

    Science.gov (United States)

    2010-04-01

    ... short-term rate prescribed by section 1274(d) for the month of the transfer and the compounding period... interest holder in excess of daily accruals. 1.860E-1 Section 1.860E-1 Internal Revenue INTERNAL REVENUE... Trusts § 1.860E-1 Treatment of taxable income of a residual interest holder in excess of daily accruals...

  6. 内生枯草芽孢杆菌E1R-J对萝卜、白菜促生作用%The Growth-promoting Effect of Endophytic Bacteria E1R-J on Radish and Cabbage

    Institute of Scientific and Technical Information of China (English)

    王心选; 高小宁; 郑刚; 王辉; 魏国荣; 康振生; 黄丽丽

    2009-01-01

    Effect of entophytic bacteria strain E1R-J on cabbage and radish seed germination, seedling growth was studied in plates, pot and outdoor field experiment. The results showed that the cell-free filtrate of strain E1R-J in different dilution and cell suspension promoted cabbage seed germination. In pot experiment, compared with untreated control, cabbage seedling height, fresh weight, dry weight increased 53% ,200% ,700% , and radish seedling height, fresh weight, dry weight increased 24. 4%, 215%, 159% after treated with 10 ×diluted E1R-J cell-free filtrate. The promotion of seedling growth by strain E1R-J was also found in the outdoor plot experiments. The chlorophyll content in the functional Leaves of the cabbage seedlings greatly increased in treatment of strain E1R-J 10×diluted cell-free filtrate and cell suspension of 10~7 cfu/mL compared with untreated control.%通过萝卜、白菜的种子萌发试验、幼苗盆栽试验以及小区试验,对小麦内生枯草芽孢杆菌E1R-J菌株的促生作用进行了研究.结果表明:E1R-J无菌滤液不同浓度稀释液及菌悬液可促进白菜种子的萌发,使萌芽整齐、萌发率增高.其无菌滤液10倍稀释液浇灌处理的盆栽白菜幼苗,株高、鲜质量、干质量分别比清水对照增长53%、200%和700%.同时发现,浇灌处理的盆栽萝卜幼苗与清水对照相比,株高、鲜质量、干质量也分别增长24.4%、215%和159%.田间小区试验结果也证明内生枯草芽孢杆菌E1R-J具有促生作用,但促生效果低于盆栽试验.利用丙酮直接浸提法测定盆栽白菜叶片中的叶绿素含量,发现无菌滤液10倍稀释液和菌悬液10倍稀释液处理的叶绿素含量增高2倍左右.

  7. The Design of Wireless Baby Monitoring System Based on nRF24E1%基于nRF24E1的无线婴儿监护系统的设计

    Institute of Scientific and Technical Information of China (English)

    马帅

    2012-01-01

    The design of a wireless baby monitoring system based on nRF24E1 is explained,the configuration of the system is introduced,and then the work theory of transmission and reception of signal is analyzed in detail.At last,the software design is described.%概述了一种基于nRF24E1的无线婴儿监护系统,介绍了该系统的总体设计结构,详细介绍了信号发送和接收的工作原理,最后介绍了软件设计部分。

  8. The Competitive Relationships of SN2, SN1, E2, E1 Reactions in Organic Chemistry%有机化学中SN2,SN1,E2,E1反应的相互竞争关系

    Institute of Scientific and Technical Information of China (English)

    颜志明; 潘英明

    2014-01-01

    有机化学中的SN2,SN1,E2,E1反应在学习中有着重要的作用,同时也是学习中易混淆的知识点.通过总结它们的反应机理、反应影响因素和反应相互竞争关系图,并利用竞争关系图解答了一些典型考研真题,方便读者理解和掌握SN2,SN1,E2,E1反应的相互竞争关系.

  9. FLYING UNITED

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Apart from selling hundreds of airplanes to China, Boeing buys locally made aircraft parts and transfers technology, in the true spirit of partnership Whenever Boeing's senior manager hear of a visit by one of China's state leaders, it's no doubt cause for celebration. Since China and the United States established diplomatic ties in 1978, every official trip by China's top statesmen has included a meeting with Boeing that

  10. Crystal structures of hydrogen-bonded co-crystals as liquid crystal precursors: 4-(n-pent-yloxy)benzoic acid-(E)-1,2-bis-(pyridin-4-yl)ethene (2/1) and 4-(n-hex-yloxy)benzoic acid-(E)-1,2-bis-(pyridin-4-yl)ethene (2/1).

    Science.gov (United States)

    Tabuchi, Yohei; Gotoh, Kazuma; Ishida, Hiroyuki

    2016-12-01

    The crystal structures of title hydrogen-bonded co-crystals, 2C12H16O3·C12H10N2, (I), and 2C13H18O3·C12H10N2, (II), have been determined at 93 K. In (I), the asymmetric unit consists of one 4-(n-pent-yloxy)benzoic acid mol-ecule and one half-mol-ecule of (E)-1,2-bis-(pyridin-4-yl)ethene, which lies about an inversion centre. The asymmetric unit of (II) comprises two crystallographically independent 4-(n-hex-yloxy)benzoic acid mol-ecules and one 1,2-bis-(pyridin-4-yl)ethene mol-ecule. In each crystal, the acid and base components are linked by O-H⋯N hydrogen bonds, forming a linear hydrogen-bonded 2:1 unit of the acid and the base. The 2:1 units are linked via C-H⋯π and π-π inter-actions [centroid-centroid distances of 3.661 (2) and 3.909 (2) Å for (I), and 3.546 (2)-3.725 (4) Å for (II)], forming column structures. In (II), the base mol-ecule is orientationally disordered over two sets of sites approximately around the N⋯N mol-ecular axis, with an occupancy ratio of 0.647 (4):0.353 (4), and the average structure of the 2:1 unit adopts nearly pseudo-C2 symmetry. Both compounds show liquid-crystal behaviour.

  11. Oxidation of Methyl and Ethyl Nitrosamines by Cytochromes P450 2E1 and 2B1

    Science.gov (United States)

    Chowdhury, Goutam; Calcutt, M. Wade; Nagy, Leslie D.; Guengerich, F. Peter

    2012-01-01

    Cytochrome P450 (P450) 2E1 is the major enzyme that oxidizes N-nitrosodimethylamine (N,N-dimethylnitrosamine, DMN), a carcinogen and also a representative of some nitrosamines formed endogenously. Oxidation of DMN by rat or human P450 2E1 to HCHO showed a high apparent intrinsic kinetic deuterium isotope effect (KIE), ≥ 8. The KIE was not attenuated in non-competitive intermolecular experiments with rat liver microsomes (DV 12.5, D(V/K) 10.9, nomenclature of Northrop, D.B. (1982) Methods Enzymol. 87, 607–625) but was with purified human P450 2E1 (DV 3.3, D(V/K) 3.7), indicating that C-H bond breaking is partially rate-limiting with human P450 2E1. With N-nitrosodiethylamine (N,N-diethylnitrosamine, DEN), the intrinsic KIE was slightly lower and was not expressed (e.g., D(V/K) 1.2) in non-competitive intermolecular experiments. The same general pattern of KIEs was also seen in the D(V/K) results with DMN and DEN for the minor products resulting from the denitrosation reactions (CH3NH2, CH3CH2NH2, and NO2−). Experiments with deuterated N-nitroso-N-methyl,N-ethylamine demonstrated that the lower KIEs associated for ethyl compared to methyl oxidation could be distinguished within a single molecule. P450 2E1 oxidized DMN and DEN to aldehydes and then to the carboxylic acids. No kinetic lags were observed in acid formation; pulse-chase experiments with carrier aldehydes showed only limited equilibration with P450 2E1-bound aldehydes, indicative of processive reactions, as reported for P450 2A6 (Chowdhury, G. et al. (2010) J. Biol. Chem. 285, 8031–8044). These same features (no lag phase for HCO2H formation, lack of equilibration in pulse-chase assays) were also seen with (rat) P450 2B1, which has lower catalytic efficiency for DMN oxidation and a larger active site. Thus, the processivity of dialkylnitrosamine oxidation appears to be shared by a number of P450s. PMID:23186213

  12. Oxidation of methyl and ethyl nitrosamines by cytochrome P450 2E1 and 2B1.

    Science.gov (United States)

    Chowdhury, Goutam; Calcutt, M Wade; Nagy, Leslie D; Guengerich, F Peter

    2012-12-18

    Cytochrome P450 (P450) 2E1 is the major enzyme that oxidizes N-nitrosodimethylamine [N,N-dimethylnitrosamine (DMN)], a carcinogen and also a representative of some nitrosamines formed endogenously. Oxidation of DMN by rat or human P450 2E1 to HCHO showed a high apparent intrinsic kinetic deuterium isotope effect (KIE), ≥8. The KIE was not attenuated in noncompetitive intermolecular experiments with rat liver microsomes {(D)V = 12.5; (D)(V/K) = 10.9 [nomenclature of Northrop, D. B. (1982) Methods Enzymol. 87, 607-625]} but was with purified human P450 2E1 [(D)V = 3.3; (D)(V/K) = 3.7], indicating that C-H bond breaking is partially rate-limiting with human P450 2E1. With N-nitrosodiethylamine [N,N-diethylnitrosamine (DEN)], the intrinsic KIE was slightly lower and was not expressed [e.g., (D)(V/K) = 1.2] in noncompetitive intermolecular experiments. The same general pattern of KIEs was also seen in the (D)(V/K) results with DMN and DEN for the minor products resulting from the denitrosation reactions (CH(3)NH(2), CH(3)CH(2)NH(2), and NO(2)(-)). Experiments with deuterated N-nitroso-N-methyl-N-ethylamine demonstrated that the lower KIEs associated with ethyl versus methyl oxidation could be distinguished within a single molecule. P450 2E1 oxidized DMN and DEN to aldehydes and then to the carboxylic acids. No kinetic lags were observed in acid formation; pulse-chase experiments with carrier aldehydes showed only limited equilibration with P450 2E1-bound aldehydes, indicative of processive reactions, as reported for P450 2A6 [Chowdhury, G., et al. (2010) J. Biol. Chem. 285, 8031-8044]. These same features (no lag phase for HCO(2)H formation and a lack of equilibration in pulse-chase assays) were also seen with (rat) P450 2B1, which has a lower catalytic efficiency for DMN oxidation and a larger active site. Thus, the processivity of dialkyl nitrosamine oxidation appears to be shared by a number of P450s.

  13. Mechanism of E1A-mediated escape from ras-induced senescence in human fibraobIasts%E1A阻断ras诱导人成纤维细胞衰老机制的研究

    Institute of Scientific and Technical Information of China (English)

    李亦蕾; 余乐

    2011-01-01

    Objective To study the effect of binding activities of the NH2 terminus of E1A to the proteins regulating cell growth on ras-induced cell senescence and explore the mechanism of ElA-mediated escape from ras-induced senescence by E1A in human fibroblast. Methods In primary human fibroblasts, the proteins regulating cell growth in association with E1A NH2 terminus, including the Rb family proteins, p300/CBP, and p400, were inactivated or interfered. The effect of alterations in the binding activities of these proteins on cell senescence bypass mediated by E1A was evaluated by cell growth curve. Results Inactivation of Rb family proteins alone was not sufficient to rescue ras-induced cell senescence, whereas inactivation of both the Rb proteins and p300/CBP blocked ras-induced senescence of human fibroblasts. Conclusion Rb and p300/CBP binding activities are both required for E1A to bypass ras-induced senescence in human fibroblasts.%目的 通过研究ElA氨基端细胞生长调节蛋白结合活性对其阻断ras诱导的细胞衰老的影响,以明确在人类成纤维细胞ElA阻断ras诱导的细胞衰老的机制.方法 采用原代培养的人类成纤维细胞,通过灭活或干扰与ElA氨基端相关的细胞生长调节蛋白,包括Rb家族蛋白、p300/CBP、p400,利用细胞生长曲线确定这些蛋白结合活性对于ElA阻断ras诱导的细胞衰老的作用.结果 单纯灭活Rb家族蛋白不能阻断ras诱导的细胞衰老,而同时灭活Rb和p300/CBP即可阻断r弱诱导的细胞衰老.结论 Rb和p300/CBP的结合活性均是EIA阻断ras诱导的人成纤维细胞细胞衰老所必需的.

  14. CYP2E1RsaⅠ/PstⅠ位点多态性与口腔癌易感性的Meta分析%A meta-analysis of CYP2E1 RsaⅠ /PstⅠ polymorphism and oral cancer risk

    Institute of Scientific and Technical Information of China (English)

    牛玉明; 沈铭; 陈盛; 陈睿; 倪小兵; 陈永吉; 胡媛媛

    2012-01-01

    Objective:To assess the relationship between CYPP4502E1 Rsa IPst I genetic polymorphism and oral cancer. Method: The relevant literatures were search in VIP, CNKI and Pubmed. The data of genotype distributions and other information were extracted in the groups of patients and healthy control and calculated with the RevMan 4.2 software for heterogeneity test and Odd rations. Result: 12 published case-control studies with 1259 patients with oral cancer and 2262 controls were selected. Overall.the pooled ORs indicated a significant association between CYP2E1 Rsa I / Pst I polymorphism and oral cancer risk (for cl / c2 vs. cl / cl (OR=I.30,95 % CI=1.04-1.62) .cl / c2+c2 / c2 vs. cl / cl (OR=1.32,95 % CI=1.07-1.64). Conclusion:This meta-analysis demonstrates that CYP2EI Rsa IPst I c2 allele may be an increase risk factor for oral cancer.%目的:运用Meta的分析方法研究CYP2E1 Rsa Ⅰ/Pst Ⅰ多态性与口腔癌易感性的相互关系.方法:检索VIP,CNKI和PubMed数据库中有关CYP2E1 Rsa Ⅰ/Pst Ⅰ多态性与口腔癌易感性关联研究的文献,以OR值和95%的可信区间为效应指标,应用Rev Man 4.2软件进行Meta分析,并使用STATA 11.0软件对发表偏倚进行检验.结果:纳入12个病例对照研究,共计1259例口腔癌患者和2262例正常对照,Meta分析结果显示,总人群中c1 /c2 vs.c1/c1(OR=1.30,95%CI=1.04~1.62),c1/c2+c2/c2 vs.c1/c1 (0R=1.32,95%CI=1.07-1.64).结论:CYP2E1突变基因型c2可能会增加口腔癌发生的风险.

  15. Genetic transformation of sweet oranges with the D4E1 gene driven by the AtPP2 promoter

    Directory of Open Access Journals (Sweden)

    Lísia Borges Attílio

    2013-07-01

    Full Text Available The objective of this work was to produce transgenic 'Pêra' and 'Valência' sweet orange plants using the D4E1 gene driven by the Arabidopsis thaliana phloem protein (AtPP2 promoter and to quantify transgene expression in different transformation events. Genetic transformation experiments were carried out with epicotyl segments co‑cultivated with Agrobacterium tumefaciens. Six plants from 'Pêra' sweet orange and seven plants from 'Valência' sweet orange were confirmed as different transgenic events by means of the polymerase chain reaction (PCR and the Southern blot techniques. Transgene expression was quantified using real‑time quantitative PCR. D4E1 gene expression levels vary from 5 up to 50 times among different transformation events.

  16. HPV18 E1^E4 is assembled into aggresome-like compartment and involved in sequestration of viral oncoproteins.

    Directory of Open Access Journals (Sweden)

    Naoko eKajitani

    2013-08-01

    Full Text Available Papillomavirus is the etiological agent for warts and several squamous carcinomas. Skin cancer induced by cottontail rabbit papillomavirus (CRPV was the first animal model for virus-induced carcinogenesis. The target organ of the virus infection is stratified epithelium and virus replication is tightly regulated by the differentiation program of the host cell. E1^E4 protein is a viral gene product, and although it is considered to be involved in the control of virus replication, little is known about the biological role. We found that HPV18 E1^E4 was assembled into an aggresome-like compartment and was involved in sequestration of virus oncoproteins, which might contribute to the differentiation-dependent lifecycle of papillomavirus.

  17. E1 strength function for two-neutron halo nuclei in an analytical three-body approach

    Energy Technology Data Exchange (ETDEWEB)

    Pushkin, A. [SENTEF, Department of Physics, University of Bergen (Norway); Jonson, B.; Zhukov, M.V. [Department of Physics, Chalmers University of Technology and University of Gothenburg, S - 412 96 Goeteborg (Sweden)

    1996-08-01

    An analytical expression for the E1 strength function for two-neutron halo nuclei is derived in a three-body model. Yukawa type wavefunctions for the ground state and three-body plane waves for the final state were used. The expression reproduces the shape and the position of the maximum of the experimental strength function for {sup 11}Li well. It is shown that the exact expression can be approximated over a large energy range with a simple function by introducing an effective two-neutron separation energy. This provides a theoretical approach for quick estimates of the E1 strength function for two-body halo nuclei. (author). Letter-to-the-editor.

  18. In-flight performance and preliminary observational results of Solar Wind Ion Detectors (SWIDs) on Chang'E-1

    Science.gov (United States)

    Kong, L. G.; Wang, S. J.; Wang, X. Y.; Zhang, A. B.; Zhu, G. W.; Yu, D. J.; Ren, Q. Y.; Reme, H.; Aoustin, C.; Zhang, X. G.; Feng, Y. B.; Zeng, L.

    2012-03-01

    SWIDs (Solar Wind Ion Detectors, SWID-A and SWID-B) are two of the scientific instruments on Chang'E-1, the first Chinese lunar mission. SWIDs utilize top-hat electrostatic analyzer to measure the low energy (alpha beam in solar wind. This is the first double ion beams event reported near the Moon. The double ion beams appeared having relatively anisotropic characteristics due to the interaction between solar wind and the Moon.

  19. Association between Rare Variants in AP4E1, a Component of Intracellular Trafficking, and Persistent Stuttering.

    Science.gov (United States)

    Raza, M Hashim; Mattera, Rafael; Morell, Robert; Sainz, Eduardo; Rahn, Rachel; Gutierrez, Joanne; Paris, Emily; Root, Jessica; Solomon, Beth; Brewer, Carmen; Basra, M Asim Raza; Khan, Shaheen; Riazuddin, Sheikh; Braun, Allen; Bonifacino, Juan S; Drayna, Dennis

    2015-11-05

    Stuttering is a common, highly heritable neurodevelopmental disorder characterized by deficits in the volitional control of speech. Whole-exome sequencing identified two heterozygous AP4E1 coding variants, c.1549G>A (p.Val517Ile) and c.2401G>A (p.Glu801Lys), that co-segregate with persistent developmental stuttering in a large Cameroonian family, and we observed the same two variants in unrelated Cameroonians with persistent stuttering. We found 23 other rare variants, including predicted loss-of-function variants, in AP4E1 in unrelated stuttering individuals in Cameroon, Pakistan, and North America. The rate of rare variants in AP4E1 was significantly higher in unrelated Pakistani and Cameroonian stuttering individuals than in population-matched control individuals, and coding variants in this gene are exceptionally rare in the general sub-Saharan West African, South Asian, and North American populations. Clinical examination of the Cameroonian family members failed to identify any symptoms previously reported in rare individuals carrying homozygous loss-of-function mutations in this gene. AP4E1 encodes the ε subunit of the heterotetrameric (ε-β4-μ4-σ4) AP-4 complex, involved in protein sorting at the trans-Golgi network. We found that the μ4 subunit of AP-4 interacts with NAGPA, an enzyme involved in the synthesis of the mannose 6-phosphate signal that targets acid hydrolases to the lysosome and the product of a gene previously associated with stuttering. These findings implicate deficits in intracellular trafficking in persistent stuttering.

  20. Synthesis, Physical Characterization, Antibacterial and Antifungal activities of a novel bis(3-((E-1-(2-hydroxyphenylethylideneaminophenylmethanone

    Directory of Open Access Journals (Sweden)

    S. Rezaei

    2006-06-01

    Full Text Available Bis (3-((E-1-(2-hydroxyphenyl ethylideneamino phenyl methanone has been synthesized in this paper and its structure was confirmed by 1H-NMR, 13C-NMR, IR and Mass spectra. Its AM1 and B3LYP/6-31G* calculations to characterize the physical properties of this molecule has been also presented. Finally, the antifungal and antibacterial activities of this derivative have been evaluated.

  1. Functional expression of 5-HT{sub 2A} receptor in osteoblastic MC3T3-E1 cells

    Energy Technology Data Exchange (ETDEWEB)

    Hirai, Takao; Kaneshige, Kota; Kurosaki, Teruko [Department of Molecular Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 1 Gakuen-cho, Fukuyama, Hiroshima 729-0292 (Japan); Nishio, Hiroaki, E-mail: nishio@fupharm.fukuyama-u.ac.jp [Department of Molecular Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 1 Gakuen-cho, Fukuyama, Hiroshima 729-0292 (Japan)

    2010-05-28

    In the previous study, we reported the gene expression for proteins related to the function of 5-hydroxytryptamine (5-HT, serotonin) and elucidated the expression patterns of 5-HT{sub 2} receptor subtypes in mouse osteoblasts. In the present study, we evaluated the possible involvement of 5-HT receptor subtypes and its inactivation system in MC3T3-E1 cells, an osteoblast cell line. DOI, a 5-HT{sub 2A} and 5-HT{sub 2C} receptor selective agonist, as well as 5-HT concentration-dependently increased proliferative activities of MC3T3-E1 cells in their premature period. This effect of 5-HT on cell proliferation were inhibited by ketanserin, a 5-HT{sub 2A} receptor specific antagonist. Moreover, both DOI-induced cell proliferation and phosphorylation of ERK1 and 2 proteins were inhibited by PD98059 and U0126, selective inhibitors of MEK in a concentration-dependent manner. Furthermore, treatment with fluoxetine, a 5-HT specific re-uptake inhibitor which inactivate the function of extracellular 5-HT, significantly increased the proliferative activities of MC3T3-E1 cells in a concentration-dependent manner. Our data indicate that 5-HT fill the role for proliferation of osteoblast cells in their premature period. Notably, 5-HT{sub 2A} receptor may be functionally expressed to regulate mechanisms underlying osteoblast cell proliferation, at least in part, through activation of ERK/MAPK pathways in MC3T3-E1 cells.

  2. Cytochrome P450 2E1 is responsible for the initiation of 1,2-dichloropropane-induced liver damage.

    Science.gov (United States)

    Yanagiba, Yukie; Suzuki, Tetsuya; Suda, Megumi; Hojo, Rieko; Gonzalez, Frank J; Nakajima, Tamie; Wang, Rui-Sheng

    2016-09-01

    1,2-Dichloropropane (1,2-DCP), a solvent, which is the main component of the cleaner used in the offset printing companies in Japan, is suspected to be the causative agent of bile duct cancer, which has been recently reported at high incidence in those offset printing workplaces. While there are some reports about the acute toxicity of 1,2-DCP, no information about its metabolism related to toxicity in animals is available. As part of our efforts toward clarifying the role of 1,2-DCP in the development of cancer, we studied the metabolic pathways and the hepatotoxic effect of 1,2-DCP in mice with or without cytochrome P450 2E1 (CYP2E1) activity. In an in vitro reaction system containing liver homogenate, 1,2-DCP was only metabolized by liver tissue of wild-type mice but not by that of cyp2e1-null mice. Furthermore, the kinetics of the solvent in mice revealed a great difference between the two genotypes; 1,2-DCP administration resulted in dose-dependent hepatic damage, as shown biochemically and pathologically, but this effect was only observed in wild-type mice. The nuclear factor κB p52 pathway was involved in the liver response to 1,2-DCP. Our results clearly indicate that the oxidative metabolism of 1,2-DCP in mice is exclusively catalyzed by CYP2E1, and this step is indispensable for the manifestation of the hepatotoxic effect of the solvent.

  3. Protective Effects of N-acetylcysteine and a Prostaglandin E1 Analog, Alprostadil, Against Hepatic Ischemia: Reperfusion Injury in Rats

    OpenAIRE

    Hsieh, Cheng-Chu; Hsieh, Shu-Chen; Chiu, Jen-Hwey; Wu, Ying-Ling

    2014-01-01

    Ischemia–reperfusion (I/R) injury has a complex pathophysiology resulting from a number of contributing factors. Therefore, it is difficult to achieve effective treatment or protection by individually targeting the mediators or mechanisms. Our aim was to analyze the individual and combined effects of N-acetylcysteine (NAC) and the prostaglandin E1 (PGE1) analog alprostadil on hepatic I/R injury in rats. Thirty male Sprague-Dawley rats were randomly divided into five groups (six rats per group...

  4. Pasteurellaceae ComE1 proteins combine the properties of fibronectin adhesins and DNA binding competence proteins.

    Directory of Open Access Journals (Sweden)

    Lisa M Mullen

    Full Text Available A novel fibronectin-binding protein from Pasteurella multocida (PM1665 that binds to the fibronectin type III(9-10 modules via two helix-hairpin-helix motifs has recently been described [1]. This protein shares homology with competence-related DNA-binding and uptake proteins (ComEA and ComE from Gram-positive and Gram-negative bacteria. Here, we show that recombinant PM1665 (now designated ComE1 also binds to DNA through the same helix-hairpin-helix motifs required for fibronectin-binding. This binding to DNA is non sequence-specific and is confined to double-stranded DNA. We have cloned and expressed ComE1 proteins from five members of the Pasteurellaceae in order to further investigate the function(s of these proteins. When expressed as recombinant GST-fusion proteins, all of the homologues bound both to fibronectin and to double-stranded DNA. Inactivation of the gene encoding the ComE1 homologue in Actinobacillus pleuropneumoniae indicates major roles for these proteins in at least two processes: natural transformation, and binding of bacteria to fibronectin.

  5. Inhibitory effect of apocynin on methylglyoxal-mediated glycation in osteoblastic MC3T3-E1 cells.

    Science.gov (United States)

    Suh, Kwang Sik; Rhee, Sang Youl; Kim, Young Seol; Choi, Eun Mi

    2015-04-01

    Methylglyoxal (MG), a highly reactive metabolite of hyperglycemia, can enhance protein glycation, oxidative stress or inflammation. The present study investigated the effects of apocynin on the mechanisms associated with MG toxicity in osteoblastic MC3T3-E1 cells. Pretreatment of MC3T3-E1 cells with apocynin prevented the MG-induced protein glycation and formation of intracellular reactive oxygen species and mitochondrial superoxide in MC3T3-E1 cells. In addition, apocynin increased glutathione levels and restored the activity of glyoxalase I inhibited by MG. These findings suggest that apocynin provide a protective action against MG-induced cell damage by reducing oxidative stress and by increasing the MG detoxification system. Apocynin treatment decreased the levels of proinflammatory cytokines such as tumor necrosis factor-α and interleukin-6 induced by MG. Additionally, the nitric oxide level reduced by MG was significantly increased by apocynin. These findings indicate that apocynin might exert its therapeutic effects via upregulation of glyoxalase system and antioxidant activity. Taken together, apocynin may prove to be an effective treatment for diabetic osteopathy.

  6. DNA Damage Reduces the Quality, but Not the Quantity of Human Papillomavirus 16 E1 and E2 DNA Replication

    Directory of Open Access Journals (Sweden)

    Molly L. Bristol

    2016-06-01

    Full Text Available Human papillomaviruses (HPVs are causative agents in almost all cervical carcinomas. HPVs are also causative agents in head and neck cancer, the cases of which are increasing rapidly. Viral replication activates the DNA damage response (DDR pathway; associated proteins are recruited to replication foci, and this pathway may serve to allow for viral genome amplification. Likewise, HPV genome double-strand breaks (DSBs could be produced during replication and could lead to linearization and viral integration. Many studies have shown that viral integration into the host genome results in unregulated expression of the viral oncogenes, E6 and E7, promoting HPV-induced carcinogenesis. Previously, we have demonstrated that DNA-damaging agents, such as etoposide, or knocking down viral replication partner proteins, such as topoisomerase II β binding protein I (TopBP1, does not reduce the level of DNA replication. Here, we investigated whether these treatments alter the quality of DNA replication by HPV16 E1 and E2. We confirm that knockdown of TopBP1 or treatment with etoposide does not reduce total levels of E1/E2-mediated DNA replication; however, the quality of replication is significantly reduced. The results demonstrate that E1 and E2 continue to replicate under genomically-stressed conditions and that this replication is mutagenic. This mutagenesis would promote the formation of substrates for integration of the viral genome into that of the host, a hallmark of cervical cancer.

  7. A mutational analysis of the ColE1-encoded cell cycle regulator Rcd confirms its role in plasmid stability.

    Science.gov (United States)

    Balding, Claire; Blaby, Ian; Summers, David

    2006-07-01

    Multimers of multicopy plasmids cause instability. They arise by homologous recombination and accumulate by over-replication in a process known as the dimer catastrophe. Dimers are resolved to monomers by site-specific recombination systems such as Xer-cer of plasmid ColE1. In addition, the Rcd checkpoint hypothesis proposes that a short transcript (Rcd) coded within ColE1 cer delays the division of multimer-containing cells. The crucial observation underpinning the checkpoint hypothesis is that when the Rcd promoter (P(cer)) is inactivated by mutation of its invariant T, the plasmid becomes unstable. Recently, we discovered that this mutation also alters a potential Fis binding site in cer. ColE1-like plasmids are less stable in fis mutant hosts and it is conceivable that instability caused by the mutation is due to altered Fis binding, rather than the loss of Rcd expression per se. We have therefore undertaken an independent test of the role of P(cer)-Rcd in multicopy plasmid stability. We have generated a series of loss-of-function mutants of Rcd and detailed analysis of two of these shows that they cause a level of instability indistinguishable from P(cer) inactivation. This result is consistent with the predictions of the checkpoint hypothesis and confirms the role of Rcd in plasmid stability.

  8. Development and evaluation of baculovirus-expressed Chikungunya virus E1 envelope proteins for serodiagnosis of Chikungunya infection.

    Science.gov (United States)

    Kumar, Pankaj; Pok, Kwoon-Yong; Tan, Li-Kiang; Angela, Chow; Leo, Yee-Sin; Ng, Lee-Ching

    2014-09-01

    Population-based serosurveillance studies provide critical estimates on community-level immunity and the potential for future outbreaks. Currently, serological assays, such as IgG enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence tests (IIFT) based on the inactivated whole virus are used to determine past Chikungunya virus (CHIKV) infection. However, these commercially available tests have variable sensitivities. To develop and evaluate recombinant based CHIKV-specific IgG antibody capture ELISAs (GAC-ELISAs), baculoviruses carrying wild-type (E1-A226, named WT) or mutant (E1-A226V, named MUT) E1 envelope protein genes of CHIKV were generated. The seroreactivity of recombinant CHIKV WT and MUT envelope proteins were determined using residual blood, collected from CHIKV-confirmed patients. The sensitivities of both recombinant CHIKV envelope proteins were 83.0% as measured by GAC-ELISAs. The specificities of both recombinant proteins were 87.8%. These GAC-ELISAs were also able to detect the persistence of anti-CHIKV IgG antibodies up to 6 months after the disease onset, together with rise in sensitivities with increasing time. These results suggest that the baculovirus purified recombinant CHIKV envelope proteins react with anti-CHIKV IgG antibodies and may be useful in population-based seroprevalence surveys. In addition, these GAC-ELISAs offer good diagnostic value to determine the recent/past CHIKV infection status in non-endemic populations.

  9. The effects of nitrogen-heme-iron coordination on substrate affinities for cytochrome P450 2E1.

    Science.gov (United States)

    Jones, Jeffrey P; Joswig-Jones, Carolyn A; Hebner, Michelle; Chu, Yuzhuo; Koop, Dennis R

    2011-08-15

    A descriptor based computational model was developed for cytochrome P450 2E1 (CYP2E1) based on inhibition constants determined for inhibition of chlorzoxazone, or 4-nitrophenol, metabolism. An empirical descriptor for type II binding was developed and tested for a series of CYP2E1 inhibitors. Inhibition constants where measured for 51 different compounds. A fast 2-dimensional predictive model was developed based on 40 compounds, and tested on 8 compounds of diverse structure. The trained model (n=40) had an r(2) value of 0.76 and an RMSE of 0.48. The correlation between the predicted and actual pK(i) values of the test set of compounds not included in the model gives an r(2) value of 0.78. The features that described binding include heme coordination (type II binding), molecular volume, octanol/water partition coefficient, solvent accessible surface area, and the sum of the atomic polarizabilities. The heme coordination parameter assigns an integer between 0 and 6 depending on structure, and is a new descriptor, based on simple quantum chemical calculations with correction for steric effects. The type II binding parameter was found to be important in obtaining a good correlation between predicted and experimental inhibition constants increasing the r(2) value from 0.38 to 0.77.

  10. Differential response of MC3T3-E1 and human mesenchymal stem cells to inositol hexakisphosphate.

    Science.gov (United States)

    Arriero, María Del Mar; Ramis, Joana M; Perelló, Joan; Monjo, Marta

    2012-01-01

    Inositol hexakisphosphate (IP6) has been found to have an important role in biomineralization. Because the complete mechanism of action of IP6 on osteoblasts is not fully understood and its potential use in the primary prevention of osteoporosis, we examined the direct effect of IP6 on cell viability and differentiation of MC3T3-E1 cells and on differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs). We show that IP6 has different effects depending on the origin of the cell target. Thus, while IP6 decreased gene expression of osteoblast markers and mineralization in MC3T3-E1 cells without negatively affecting cell viability and ALP activity, an increase in gene expression of ALP was observed in hUC-MSCs committed to the osteoblastic lineage. This increasing effect of IP6 on ALP mRNA expression levels was reversed by the addition of a selective inhibitor of IP6 kinase, suggesting that the effect of IP6 might be due through its pyrophosphorylated derivatives. Besides, Rankl mRNA levels were decreased after IP6 treatment in MC3T3-E1 cells, pointing to a paracrine effect on osteoclasts. Our results indicate that IP6 has different effects on osteoblast differentiation depending on the cell type and origin. However, further studies are needed to examine the net effect of IP6 on bone formation and its potential as novel antiosteoporosis drug. Copyright © 2012 S. Karger AG, Basel.

  11. Contributions of caspase-8 and -9 to liver injury from CYP2E1-produced metabolites of halogenated hydrocarbons.

    Science.gov (United States)

    Ijiri, Yoshio; Kato, Ryuji; Sadamatsu, Maiko; Takano, Mina; Yasuda, Yuki; Tanaka, Fumiaki; Oishi, Chiyo; Imano, Hideki; Okada, Yoshikatsu; Tanaka, Kazuhiko; Hayashi, Tetsuya

    2017-01-12

    1. Drug-induced liver injury is difficult to predict at the pre-clinical stage. This study aimed to clarify the roles of caspase-8 and -9 in CYP2E1 metabolite-induced liver injury in both rats and cell cultures in vitro treated with carbon tetrachloride (CCl4), halothane or sevoflurane. The human hepatocarcinoma functional liver cell line was maintained in 3-dimensional culture alone or in co-culture with human acute monocytic leukemia cells. 2. In vivo, laboratory indices of liver dysfunction and histology were normal after administration of sevoflurane. CCl4 treatment increased blood AST/ALT levels, liver caspase-3 and -9 activities and liver malondialdehyde, accompanied by centrilobular hepatocyte necrosis. Halothane increased AST/ALT levels, caspase-3 and -8 activities (but not malondialdehyde) concomitant with widespread hepatotoxicity. In vitro, CCl4 treatment increased caspase-9 activity and decreased both mitochondrial membrane potential (MMP) and cell viability. In co-culture, halothane increased caspase-8 activity and decreased MMP and cellular viability. There were no toxic responses in CYP2E1 knockdown in monoculture and co-culture. 3. CYP2E1-inducing compounds play a pivotal role in halogenated hydrocarbon toxicity. 4. Changes in hepatocyte caspase-8 and -9 activities could be novel biomarkers of metabolites causing DILI, and in pre-clinical development of new pharmaceuticals can predict nascent DILI in the clinical stage.

  12. STABILITAS EPIGALOKATEKIN GALAT DALAM KRIM EKSTRAK TEH HIJAU DENGAN VARIASI KONSENTRASI ANTIOKSIDAN VITAMIN C 1% DAN VITAMIN E 1%

    Directory of Open Access Journals (Sweden)

    Nining Sugihartini

    2016-11-01

    Full Text Available Epigallocatechingallate (EGCG in green tea extract has activity as an anti-inflammatory agent. On the other hand the stability of EGCG is poor because of the oxidation. The aim of this study was to determine the influence of Vitamine C and Vitamine E in formulation of green tea extract cream to the stabiliy of EGCG. The green tea extract was obtained from the extraction process by infundation followed by fractination with ethyl acetate as the solvent. The three formulas were compiled in similar composition with the concentration of vitamine C 1% (FI, Vitamine E 1% (FII and there was no Vitamine C and Vitamine E (FIII as a control. The EGCG level was determinated by TLC-densitometry methode. The stability parameter was determinated by calculated of the Q10 of each formula. The result of this study showed that the parameter of t90 of EGCG with Vitamine C 1%, Vitamine E 1% and control addition were 0.0108 hours, 0.0087 hours, 0.0084 hours, respectively. Stability of EGCG in green tea leaf extract cream with addition of the vitamin C 1% was higher than it stability with the addition of vitamin E 1%. The concentration of Vitamin C 1% was the optimum concentration as antioxidant in formulation of green tea extract cream.

  13. Rapamycin/sodium hyaluronate binding on nano-hydroxyapatite coated titanium surface improves MC3T3-E1 osteogenesis

    Science.gov (United States)

    Liu, Chao; Dong, Jian Yong; Yue, Lin Lin; Liu, Shao Hua; Wan, Yi; Liu, Hong; Tan, Wan Ye; Guo, Qian Qian; Zhang, Dong

    2017-01-01

    Endosseous titanium (Ti) implant failure due to poor biocompatibility of implant surface remains a major problem for osseointegration. Improving the topography of Ti surface may enhance osseointegration, however, the mechanism remains unknown. To investigate the effect of modified Ti surface on osteogenesis, we loaded rapamycin (RA) onto nano-hydroxyapatite (HAp) coated Ti surface which was acid-etched, alkali-heated and HAp coated sequentially. Sodium hyaluronate (SH) was employed as an intermediate layer for the load of RA, and a steady release rate of RA was maintained. Cell vitality of MC3T3-E1 was assessed by MTT. Osteogenesis of MC3T3-E1 on this modified Ti surface was evaluated by alkaline phosphatase (ALP) activity, mineralization and related osteogenesis genes osteocalcin (OCN), osteopontin (OPN), Collagen-I and Runx2. The result revealed that RA/SH-loaded nano-HAp Ti surface was innocent for cell vitality and even more beneficial for cell osteogenesis in vitro. Furthermore, osteogenesis of MC3T3-E1 showed significant association with the mammalian target of rapamycin (mTOR) phosphorylation by RA, which required further study about the mechanism. The approach to this modified Ti surface presented in this paper has high research value for the development of Ti-based implant. PMID:28182765

  14. United States

    Directory of Open Access Journals (Sweden)

    Stephen Bernow

    1998-12-01

    Full Text Available This paper presents and discusses an integrated set of policies designed to reduce U.S. carbon emissions over the next four decades. This innovation path also aims to promote environmental quality, particularly by reducing emissions of criteria air pollutants, to reduce U.S. dependence on imported oil, and to induce technological innovation and diffusion in energy production and consumption. The innovation path would reduce economy-wide carbon emissions by 26% below baseline projections for 2010 and by 62% below baseline projections for 2030; this translates into 10% below 1990 levels in 2010 and 45% below 1990 levels in 2030. Emissions of criteria pollutants also would be significantly reduced, as would petroleum imports by the United States. Moreover, the innovation path would yield cumulative net savings for the United States of $218 billion (1993 dollars through 2010, or $19 billion on a leveled annual basis, and would result in 800,000 additional jobs nationwide by 2010. Although the overall findings from the innovation path analysis are robust, the results should be taken as indicative, rather than precisely predictive, owing to uncertainties in future costs, prices, technology performance, and consumer behavior.

  15. Optimization of Acidothermus Celluloyticus Endoglucanase (E1) Production in Transgenic Tobacco Plants by Transcriptional, Post-transcription and Post-Translational Modification

    Energy Technology Data Exchange (ETDEWEB)

    Dai, Ziyu; Hooker, Brian S.; Quesenberry, Ryan D.; Thomas, S. R.

    2005-10-01

    Biochemical characteristics of Acidothermus cellulolyticus endoglucanase (E1) and its physiological effects in transgenic tobacco (Nicotiana tabacum) has been studied previously. In an attempt to obtain a high level of production of intact E1 in transgenic plants, the E1 gene was expressed under the control of strong Mac promoter (a hybrid promoter of manopine synthase promoter and cauliflower mosaic virus 35S promoter enhancer region) or tomato Rubisco small subunit (RbcS-3C) promoter with different 5’ untranslated leader (UTL) sequence and targeted to different subcellular comartmentations with various transit peptides. The expression of E1 protein in transgenic tobacco plants was determined via E1 activity, protein immunobloting, and RNA gel-blotting analyses. Effects of different transit peptides on E1 protein production and its stability were examined in transgenic tobacco plants carrying one of six transgene expression vectors with the same (Mac) promoter and transcription terminator (Tmas). Transgenic tobacco plants with apoplast transit peptide (Mm-apo) had the highest average E1 activity and protein accumulation , while E1 protein was more stable in transgenic plants with no transit peptide (Mm) than others. The E1 expression under tomato RbcS-3C promoter was higher than that under Mac promoter based on the average E1 activity, E1 protein accumulation, and RNA gel-blotting. The E1 expression was increased more than two fold when the 5’-UTL of alfalfa mosaic virus RNA4 gene replaced the UTL of RbcS-3C promoter, while the UTL of alfalfa mosaic virus RNA4 gene was less effective than the UTL of Mac promoter. The optimal combination of promoter, 5’-UTL, and subcellular compartmentation (transit peptide) for E1 protein production in transgenic tobacco plants are discussed.

  16. Prostaglandin E1 joint plasma exchange treatment of chronic severe hepatitis%前列腺素E1联合血浆置换治疗慢性重型肝炎的临床研究

    Institute of Scientific and Technical Information of China (English)

    唐学兰

    2011-01-01

    目的 探讨前列腺素E1联合血浆置换治疗慢性重型肝炎的疗效.方法 将88例慢性重型肝炎患者完全随机分为治疗组与对照组.治疗组46例,在内科综合治疗的基础上予前列腺素E1联合血浆置换治疗;对照组42例,只予内科综合治疗.观察2组治疗前后的症状体征及相关实验室指标.结果 治疗组经治疗后症状体征均明显好转,总胆红素、ALT、总蛋白、白蛋白、胆碱酯酶、凝血酶原活动度、血氨均改善明显(P<0.01或P<0.05).治疗后,2组间除白蛋白水平外,其余各指标均差异有统计学意义(P<0.05).随访3个月,治疗组总有效率69.6%(32例)、早期有效率79.3%(23例)、中期有效率64.3%(9例),均明显高于对照组的47.6%(20例)、55.6%(15例)、38.5%(5例),差异均有统计学意义(均P<0.01).结论 前列腺素E1联合血浆置换治疗慢性重型肝炎能明显改善肝功能,明显提高疗效.%Objective To investigate the curative effect, timing and feasibility of prostaglandin E1 combined plasma exchange in treatment of chronic severe hepatitis. Methods Eighty-eight patients with chronic severe hepatitis were randomly divided into treatment group and control group. The treatment group was given prostaglandin E1 combined plasma exchange treatment based on the comprehensive treatment of internal medicine. Control group only had given the internal medicine comprehensive treatment. Results The symptoms and signs of the treatment group were improved after treatments, especially with total bilirubin, cholinesterase, prothrombin activity, NH3 ( P < 0.01,P < 0.05 ). After treatment, except for albumin, the others indicators of the two groups had significant difference ( P < 0.05). Follow-up for three months, the total effective rate of the treatment group was 69.6% ( 32cases ), early effective rate was 79.3% (23 cases), mid-term effective rate was 64.3% (9 cases) were all significantly higher than those of control group [47

  17. 前列腺素E-1对过度膨胀犬肺血流动力学的影响%Effect of Prostaglandin E-1 on Hemodynamics of Hyperinflation Lung in Canine

    Institute of Scientific and Technical Information of China (English)

    张璟; 黄韬; 洪志鹏; 尹小川; 段晋; 赵炜; 李定彪; 杨学庆

    2007-01-01

    [目的]探讨过度膨胀对肺血流动力学的影响以及前列腺素E-1的保护作用.[方法]杂种犬16只,随机分为对照组和实验组,建立肺过度膨胀动物模型.对照组:保持肺过度膨胀6 h;实验组:保持肺过度膨胀6h,并以30 ng/(kg·min)的剂量给予前列腺素E-1,两组6 h内均进行连续的肺动脉压、体循环动脉压及中心静脉压的监测,6 h后取标本进行血栓素B2的检测.[结果]两组肺动脉压、体循环动脉压及中心静脉压术前差异均无显著性(P>0.05),术后两组肺动脉压都有升高,对照组明显高于实验组(P<0.05),两组体循环动脉压及中心静脉压变化差异无显著性(P>0.05),对照组血栓索B2明显高于实验组(P<0.05);.[结论]前列腺素E-1可改善过度膨胀所致的犬肺动脉高压,其作用机制与抑制TxA2合成,纠正TXA2/PGI2比值失调等有关.

  18. Annealing mechanisms of self-interstitial related defect E1=E{sub c}-0.39 eV in irradiated silicon

    Energy Technology Data Exchange (ETDEWEB)

    Abdullin, Kh.A., E-mail: kh.a.abdullin@mail.ru [National Nanotechnology Open Laboratory, al-Farabi Kazakh National University, 71 al-Farabi Str., 050060 Almaty (Kazakhstan); Mukashev, B.N. [National Nanotechnology Open Laboratory, al-Farabi Kazakh National University, 71 al-Farabi Str., 050060 Almaty (Kazakhstan)

    2012-07-01

    DLTS and thermally stimulated capacitance (TSCap) studies of {alpha}-particle irradiated p-Si were undertaken to obtain additional information about the self-interstitial related defect E1=E{sub c}-0.39 eV. The E1 defect can be retained frozen up to room temperature without any minority carrier injection, but under injection conditions at 77-300 K the E1 becomes mobile. As a result of annealing of the E1 defect, the carbon interstitial concentration grows. The E1 defect production rate under reverse bias as well as thermal annealing behavior depends strongly on the temperature and impurity concentration. Numerical estimates of the E1 migration via a Bourgoin-Corbett mechanism are in good agreement with the experimental data obtained.

  19. 基于 E1椭球的 GPS 二维约束网平差设计与实现%The Design and Implementation Of GPS 2D Constrained Net Adjustment Based on the E1 Ellipsoid

    Institute of Scientific and Technical Information of China (English)

    杨剑伟; 吕志伟; 白文礼; 吕金浩

    2013-01-01

    现代工程G PS基线向量网与地面网联合平差一般是采用二维约束平差的方式在平面直角坐标系中进行。本文详述了基于E1椭球在地方独立坐标系中进行二维约束网平差的原理。算例表明:对于小范围的G PS工程控制网,采用二维约束平差在精度和实用性方面都能满足需求。%Modern engineering of GPS baseline vector network and the ground network combined adjustment is generally conducted based on the 2D constrained adjustment manner in the plane rectangular coordinate system .2D constrained net adjustment is described based on E1 ellipsoid in local independent coordinate system detailed .The example shows that in the small range of GPS control network ,the 2D constrained network adjustment can meet the demand in terms of the accuracy and practicality .

  20. Dissemination and transmission of the E1-226V variant of chikungunya virus in Aedes albopictus are controlled at the midgut barrier level.

    Directory of Open Access Journals (Sweden)

    Camilo Arias-Goeta

    Full Text Available Emergence of arboviruses could result from their ability to exploit new environments, for example a new host. This ability is facilitated by the high mutation rate occurring during viral genome replication. The last emergence of chikungunya in the Indian Ocean region corroborates this statement since a single viral mutation at the position 226 on the E1 glycoprotein (E1-A226V was associated with enhanced transmission by the mosquito Aedes albopictus in regions where the major mosquito vector, Aedes aegypti, is absent.We used direct competition assays in vivo to dissect out the mechanisms underlying the selection of E1-226V by Ae. albopictus. When the original variant E1-226A and the newly emerged E1-226V were provided in the same blood-meal at equal titers to both species of mosquitoes, we found that the proportion of both variants was drastically different in the two mosquito species. Following ingestion of the infectious blood-meal, the E1-226V variant was preferentially selected in Ae. albopictus, whereas the E1-226A variant was sometimes favored in Ae. aegypti. Interestingly, when the two variants were introduced into the mosquitoes by intrathoracic inoculations, E1-226V was no longer favored for dissemination and transmission in Ae. albopictus, showing that the midgut barrier plays a key role in E1-226V selection.This study sheds light on the role of the midgut barrier in the selection of novel arbovirus emerging variants. We also bring new insight into how the pre-existing variant E1-226V was selected among other viral variants including E1-226A. Indeed the E1-226V variant present at low levels in natural viral populations could rapidly emerge after being selected in Ae. albopictus at the midgut barrier level.

  1. 26 CFR 1.404(e)-1A - Contributions on behalf of a self-employed individual to or under a qualified pension, annuity...

    Science.gov (United States)

    2010-04-01

    ... individual to or under a qualified pension, annuity, or profit-sharing plan. 1.404(e)-1A Section 1.404(e)-1A...) INCOME TAXES Pension, Profit-Sharing, Stock Bonus Plans, Etc. § 1.404(e)-1A Contributions on behalf of a self-employed individual to or under a qualified pension, annuity, or profit-sharing plan. (a) In...

  2. Sulfation of fulvestrant by human liver cytosols and recombinant SULT1A1 and SULT1E1

    Directory of Open Access Journals (Sweden)

    Edavana VK

    2011-11-01

    Full Text Available Vineetha Koroth Edavana1, Xinfeng Yu1, Ishwori B Dhakal1, Suzanne Williams1, Baitang Ning2, Ian T Cook3, David Caldwell1, Charles N Falany3, Susan Kadlubar11Division of Medical Genetics, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, AR, USA; 2Division of Personalized Nutrition and Medicine, National Center for Toxicological Research, Food and Drug Administration, Jefferson, AR, USA; 3Department of Pharmacology, University of Alabama, Birmingham, AL, USAAbstract: Fulvestrant (Faslodex™ is a pure antiestrogen that is approved to treat hormone receptor-positive metastatic breast cancer in postmenopausal women. Previous studies have demonstrated that fulvestrant metabolism in humans involves cytochromes P450 and UDP-glucuronosyltransferases (UGTs. To date, fulvestrant sulfation has not been characterized. This study examined fulvestrant sulfation with nine recombinant sulfotransferases and found that only SULT1A1 and SULT1E1 displayed catalytic activity toward this substrate, with Km of 4.2 ± 0.99 and 0.2 ± 0.16 µM, respectively. In vitro assays of 104 human liver cytosols revealed marked individual variability that was highly correlated with β-naphthol sulfation (SULT1A1 diagnostic substrate; r = 0.98, P < 0.0001, but not with 17ß-estradiol sulfation (SULT1E1 diagnostic substrate; r = 0.16, P = 0.10. Fulvestrant sulfation was correlated with both SULT1A1*1/2 genotype (P value = 0.023 and copy number (P < 0.0001. These studies suggest that factors influencing SULT1A1/1E1 tissue expression and/or enzymatic activity could influence the efficacy of fulvestrant therapy.Keywords: fulvestrant, sulfotransferase, genotype, copy number

  3. Attenuation of microRNA-16 derepresses the cyclins D1, D2 and E1 to provoke cardiomyocyte hypertrophy

    Science.gov (United States)

    Huang, Shuai; Zou, Xiao; Zhu, Jie-Ning; Fu, Yong-Heng; Lin, Qiu-Xiong; Liang, Ye-You; Deng, Chun-Yu; Kuang, Su-Juan; Zhang, Meng-Zhen; Liao, Yu-Lin; Zheng, Xi-Long; Yu, Xi-Yong; Shan, Zhi-Xin

    2015-01-01

    Cyclins/retinoblastoma protein (pRb) pathway participates in cardiomyocyte hypertrophy. MicroRNAs (miRNAs), the endogenous small non-coding RNAs, were recognized to play significant roles in cardiac hypertrophy. But, it remains unknown whether cyclin/Rb pathway is modulated by miRNAs during cardiac hypertrophy. This study investigates the potential role of microRNA-16 (miR-16) in modulating cyclin/Rb pathway during cardiomyocyte hypertrophy. An animal model of hypertrophy was established in a rat with abdominal aortic constriction (AAC), and in a mouse with transverse aortic constriction (TAC) and in a mouse with subcutaneous injection of phenylephrine (PE) respectively. In addition, a cell model of hypertrophy was also achieved based on PE-promoted neonatal rat ventricular cardiomyocyte and based on Ang-II-induced neonatal mouse ventricular cardiomyocyte respectively. We demonstrated that miR-16 expression was markedly decreased in hypertrophic myocardium and hypertrophic cardiomyocytes in rats and mice. Overexpression of miR-16 suppressed rat cardiac hypertrophy and hypertrophic phenotype of cultured cardiomyocytes, and inhibition of miR-16 induced a hypertrophic phenotype in cardiomyocytes. Expressions of cyclins D1, D2 and E1, and the phosphorylated pRb were increased in hypertrophic myocardium and hypertrophic cardiomyocytes, but could be reversed by enforced expression of miR-16. Cyclins D1, D2 and E1, not pRb, were further validated to be modulated post-transcriptionally by miR-16. In addition, the signal transducer and activator of transcription-3 and c-Myc were activated during myocardial hypertrophy, and inhibitions of them prevented miR-16 attenuation. Therefore, attenuation of miR-16 provoke cardiomyocyte hypertrophy via derepressing the cyclins D1, D2 and E1, and activating cyclin/Rb pathway, revealing that miR-16 might be a target to manage cardiac hypertrophy. PMID:25583328

  4. Role of cleavage at the core-E1 junction of hepatitis C virus polyprotein in viral morphogenesis

    Science.gov (United States)

    Pène, Véronique; Lemasson, Matthieu; Harper, Francis; Pierron, Gérard; Rosenberg, Arielle R.

    2017-01-01

    In hepatitis C virus (HCV) polyprotein sequence, core protein terminates with E1 envelope signal peptide. Cleavage by signal peptidase (SP) separates E1 from the complete form of core protein, anchored in the endoplasmic reticulum (ER) membrane by the signal peptide. Subsequent cleavage of the signal peptide by signal-peptide peptidase (SPP) releases the mature form of core protein, which preferentially relocates to lipid droplets. Both of these cleavages are required for the HCV infectious cycle, supporting the idea that HCV assembly begins at the surface of lipid droplets, yet SPP-catalyzed cleavage is dispensable for initiation of budding in the ER. Here we have addressed at what step(s) of the HCV infectious cycle SP-catalyzed cleavage at the core-E1 junction is required. Taking advantage of the sole system that has allowed visualization of HCV budding events in the ER lumen of mammalian cells, we showed that, unexpectedly, mutations abolishing this cleavage did not prevent but instead tended to promote the initiation of viral budding. Moreover, even though no viral particles were released from Huh-7 cells transfected with a full-length HCV genome bearing these mutations, intracellular viral particles containing core protein protected by a membrane envelope were formed. These were visualized by electron microscopy as capsid-containing particles with a diameter of about 70 nm and 40 nm before and after delipidation, respectively, comparable to intracellular wild-type particle precursors except that they were non-infectious. Thus, our results show that SP-catalyzed cleavage is dispensable for HCV budding per se, but is required for the viral particles to acquire their infectivity and secretion. These data support the idea that HCV assembly occurs in concert with budding at the ER membrane. Furthermore, capsid-containing particles did not accumulate in the absence of SP-catalyzed cleavage, suggesting the quality of newly formed viral particles is controlled before

  5. (E-1-Phenylethanone semicarbazone

    Directory of Open Access Journals (Sweden)

    Hoong-Kun Fun

    2009-08-01

    Full Text Available In the title compound, C9H11N3O, the benzene ring is disordered over two positions with refined occupancies of 0.922 (5 and 0.078 (5. The program PLATON [Spek (2009. Acta Cryst. D65, 148–155] recommends the solution in the space group C2/m with a = 7.3050 (3, b = 6.6745 (2, c = 18.3853 (6 Å and β = 96.986 (2°. However, the large number of non-extinct reflections needed to be ignored if C2/m is chosen suggested that the space group is incorrect, even though the R values are lower than that for P21/c. The semicarbazone group is essentially planar, with a maximum deviation of 0.046 (1 Å for one of the N atoms. The mean plane of the semicarbazone group forms dihedral angles of 33.61 (8 and 39.1 (9° with the benzene ring of the major and minor components, respectively. In the crystal structure, molecules are linked by intermolecular N—H...O hydrogen bonds into extended chains along the c axis. The crystal structure is further stabilized by weak intermolucular C—H...π interactions.

  6. The Pharmacological Effects of Morroniside and Loganin Isolated from Liuweidihuang Wan, on MC3T3-E1 Cells

    OpenAIRE

    Xijun Wang; Hui Sun; Ping Wang; Kun Yang; Wei Wang; Manyu Li

    2010-01-01

    Liuweidihuang Wan (LW), initially a well-known formula for curing “wu chi wu ruan”, is commonly used nowadays for clinical treatment of Postmenopausal Osteoporosis (PO), but the identity of the effective substance(s) remains unclear. The present study was designed to evaluate the effects of morroniside and loganin isolated from LW on the proliferation, differentiation and apoptosis of MC3T3-E1 cells, as well as the possible mechanism of action. Morroniside and loganin had no effects on the pr...

  7. Cyclin-dependent kinase 2 is an ideal target for ovary tumors with elevated cyclin E1 expression

    OpenAIRE

    Yang, Liu; Fang, Dongdong; Chen, Huijun; Lu, Yiyu; Dong, Zheng; Ding, Han-Fei; Jing, Qing; Su, Shi-bing; Huang, Shuang

    2015-01-01

    CCNE1 gene amplification is present in 15-20% ovary tumor specimens. Here, we showed that Cyclin E1 (CCNE1) was overexpressed in 30% of established ovarian cancer cell lines. We also showed that CCNE1 was stained positive in over 40% of primary ovary tumor specimens regardless of their histological types while CCNE1 staining was either negative or low in normal ovary and benign ovary tumor tissues. However, the status of CCNE1 overexpression was not associated with the tumorigenic potential o...

  8. Coactivator-associated arginine methyltransferase 1 (CARM1) is a positive regulator of the Cyclin E1 gene

    OpenAIRE

    El Messaoudi, Selma; Fabbrizio, Eric; Rodriguez, Carmen; Chuchana, Paul; Fauquier, Lucas; Cheng, Donghang; Theillet, Charles; Vandel, Laurence; Bedford, Mark T.; Sardet, Claude

    2006-01-01

    The Cyclin E1 gene (CCNE1) is an ideal model to explore the mechanisms that control the transcription of cell cycle-regulated genes whose expression rises transiently before entry into S phase. E2F-dependent regulation of the CCNE1 promoter was shown to correlate with changes in the level of H3-K9 acetylation/methylation of nucleosomal histones positioned at the transcriptional start site region. Here we show that, upon growth stimulation, the same region is subject to variations of H3-R17 an...

  9. Sequestration of p53 in the Cytoplasm by Adenovirus Type 12 E1B 55-Kilodalton Oncoprotein Is Required for Inhibition of p53-Mediated Apoptosis

    OpenAIRE

    2003-01-01

    The adenovirus E1B 55-kDa protein is a potent inhibitor of p53-mediated transactivation and apoptosis. The proposed mechanisms include tethering the E1B repression domain to p53-responsive promoters via direct E1B-p53 interaction. Cytoplasmic sequestration of p53 by the 55-kDa protein would impose additional inhibition on p53-mediated effects. To investigate further the role of cytoplasmic sequestration of p53 in its inhibition by the E1B 55-kDa protein we systematically examined domains in b...

  10. The role of reactive oxygen species (ROS and cytochrome P-450 2E1 in the generation of carcinogenic etheno-DNA adducts

    Directory of Open Access Journals (Sweden)

    Kirsten Linhart

    2014-01-01

    Full Text Available Exocyclic etheno-DNA adducts are mutagenic and carcinogenic and are formed by the reaction of lipidperoxidation (LPO products such as 4-hydoxynonenal or malondialdehyde with DNA bases. LPO products are generated either via inflammation driven oxidative stress or via the induction of cytochrome P-450 2E1 (CYP2E1. In the liver CYP2E1 is induced by various compounds including free fatty acids, acetone and ethanol. Increased levels of CYP2E1 and thus, oxidative stress are observed in the liver of patients with non-alcoholic steatohepatitis (NASH as well as in the chronic alcoholic. In addition, chronic ethanol ingestion also increases CYP2E1 in the mucosa of the oesophagus and colon. In all these tissues CYP2E1 correlates significantly with the levels of carcinogenic etheno-DNA adducts. In contrast, in patients with non-alcoholic steatohepatitis (NASH hepatic etheno-DNA adducts do not correlate with CYP2E1 indicating that in NASH etheno-DNA adducts formation is predominately driven by inflammation rather than by CYP2E1 induction. Since etheno-DNA adducts are strong mutagens producing various types of base pair substitution mutations as well as other types of genetic damage, it is strongly believed that they are involved in ethanol mediated carcinogenesis primarily driven by the induction of CYP2E1.

  11. Characterization of Autoantibodies against the E1 Subunit of Branched-Chain 2-Oxoacid Dehydrogenase in Patients with Primary Biliary Cirrhosis

    Directory of Open Access Journals (Sweden)

    Tsutomu Mori

    2012-01-01

    Full Text Available Primary biliary cirrhosis (PBC is characterized by antimitochondrial antibodies (AMAs that react with the lipoyl-containing E2 subunits of 2-oxoacid dehydrogenase complexes such as BCOADC and PDC. The lipoyl domains of E2 contain the major epitopes essential for immunopathology. However, the non-lipoyl-containing E1 subunits are also frequently targeted. Since anti-E1 antibodies always appear in combination with anti-E2 antibodies, the mechanisms underlying the autoimmunity against E1 may be linked to, but distinct from, those against E2. Here, we demonstrate that intermolecular and intramolecular determinant spreading underlies the autoimmunity against E1. We performed characterizations and epitope mapping for anti-BCOADC-E1 antibodies from both the intermolecular and intramolecular points of view. The antibody reactivities form a cluster against the BCOADC complex that is distinct from that against the PDC complex, and the anti-BCOADC-E1 antibodies arise as part of the cluster against the BCOADC complex. Multiple epitopes are present on the surface of the BCOADC-E1 molecule, and the major epitope overlaps with the active center. Sera with anti-BCOADC-E1 antibodies strongly inhibited the enzyme activity. These findings suggest that the E1 subunit as part of the native BCOADC complex is an immunogen, and that determinant spreading is involved in the pathogenesis of AMA production.

  12. Termination unit

    Energy Technology Data Exchange (ETDEWEB)

    Traeholt, Chresten; Willen, Dag; Roden, Mark; Tolbert, Jerry C.; Lindsay, David; Fisher, Paul W.; Nielsen, Carsten Thidemann

    2016-05-03

    Cable end section comprises end-parts of N electrical phases/neutral, and a thermally-insulation envelope comprising cooling fluid. The end-parts each comprises a conductor and are arranged with phase 1 innermost, N outermost surrounded by the neutral, electrical insulation being between phases and N and neutral. The end-parts comprise contacting surfaces located sequentially along the longitudinal extension of the end-section. A termination unit has an insulating envelope connected to a cryostat, special parts at both ends comprising an adapter piece at the cable interface and a closing end-piece terminating the envelope in the end-section. The special parts houses an inlet and/or outlet for cooling fluid. The space between an inner wall of the envelope and a central opening of the cable is filled with cooling fluid. The special part at the end connecting to the cryostat houses an inlet or outlet, splitting cooling flow into cable annular flow and termination annular flow.

  13. Structural comparison of cytochromes P450 2A6, 2A13, and 2E1 with pilocarpine

    Energy Technology Data Exchange (ETDEWEB)

    DeVore, Natasha M.; Meneely, Kathleen M.; Bart, Aaron G.; Stephens, Eva S.; Battaile, Kevin P.; Scott, Emily E. (Kansas); (HWMRI)

    2013-11-20

    Human xenobiotic-metabolizing cytochrome P450 (CYP) enzymes can each bind and monooxygenate a diverse set of substrates, including drugs, often producing a variety of metabolites. Additionally, a single ligand can interact with multiple CYP enzymes, but often the protein structural similarities and differences that mediate such overlapping selectivity are not well understood. Even though the CYP superfamily has a highly canonical global protein fold, there are large variations in the active site size, topology, and conformational flexibility. We have determined how a related set of three human CYP enzymes bind and interact with a common inhibitor, the muscarinic receptor agonist drug pilocarpine. Pilocarpine binds and inhibits the hepatic CYP2A6 and respiratory CYP2A13 enzymes much more efficiently than the hepatic CYP2E1 enzyme. To elucidate key residues involved in pilocarpine binding, crystal structures of CYP2A6 (2.4 {angstrom}), CYP2A13 (3.0 {angstrom}), CYP2E1 (2.35 {angstrom}), and the CYP2A6 mutant enzyme, CYP2A6 I208S/I300F/G301A/S369G (2.1 {angstrom}) have been determined with pilocarpine in the active site. In all four structures, pilocarpine coordinates to the heme iron, but comparisons reveal how individual residues lining the active sites of these three distinct human enzymes interact differently with the inhibitor pilocarpine.

  14. Ethanolic extract of Actaea racemosa (black cohosh) potentiates bone nodule formation in MC3T3-E1 preosteoblast cells.

    Science.gov (United States)

    Chan, B Y; Lau, K S; Jiang, B; Kennelly, E J; Kronenberg, F; Kung, A W C

    2008-09-01

    Aceaea racemosa (formerly Cimicifuga racemosa, black cohosh, AR) extracts have been widely used as an alternative to hormonal replacement therapy for menopausal symptoms. Recent evidences suggest AR extracts are also effective in protecting against postmenopausal bone loss. To determine whether AR has any direct anabolic effect on osteoblasts, we investigated the ethanolic extract of AR on bone nodule formation in mouse MC3T3-E1 preosteoblast cells. AR did not stimulate osteoblast proliferation. Rather, at high doses of 1000 ng/mL for 48 h, AR suppressed (7.2+/-0.9% vs. control) osteoblast proliferation. At 500 ng/mL, a significant increase in bone nodule formation was seen with Von Kossa staining. Using quantitative PCR analysis, AR was shown to enhance the gene expression of runx2 and osteocalcin. Co-treatment with ICI 182,780, the selective estrogen receptor antagonist, abolished the stimulatory effect of AR on runx2 and osteocalcin gene induction, as well as on bone nodule formation in MC3T3-E1 cells. This is a first report of the direct effect of AR on enhancement of bone nodule formation in osteoblasts, and this action was mediated via an estrogen receptor-dependent mechanism. The results provide a scientific rationale at the molecular level for the claim that AR can offer effective prevention of postmenopausal bone loss.

  15. Effects of cholesterol-bearing pullulan (CHP)-nanogels in combination with prostaglandin E1 on wound healing.

    Science.gov (United States)

    Kobayashi, Hiroshi; Katakura, Osamu; Morimoto, Nobuyuki; Akiyoshi, Kazunari; Kasugai, Shohei

    2009-10-01

    The cholesterol-bearing pullulan (CHP)-nanogels are able to trap hydrophobic drugs or proteins inside the nanogels, which is potential in application to drug delivery system and tissue engineering. On the other hand, prostaglandin E1 (PGE1) plays important roles in wound healing and PGE1 ointment has been clinically used to treat chronic skin ulcers and wounds. The purpose of this study is to evaluate effects of CHP nanogels in combination with prostaglandin E1 on wound healing in full thickness skin defect model. A square skin defect (1 x 1 cm(2)) of full thickness was created on the dorsal of Wistar rats. The wound was treated with CHP nanogels without PGE1 (CHP group) or CHP nanogels containing with PGE1 (CHP/PGE1 group) or PGE1 ointment (PGE1 ointment group). In both CHP/PGE1 and PGE1 ointment groups, approximately 6 microg of PGE1 was applied to each wound. In the control group, the wound was untreated. The wound was evaluated in measuring wound area and histologically. In CHP/PGE1 group, the rate of wound size reduction was significantly higher than the ones of other groups. Histologically, CHP/PGE1 promoted neoepithelialization, neovascularization, and wound closure compared to the other treatments. These results suggest that CHP in combination with PGE1 can promote wound healing, which confirms the efficiency of CHP nanogels-based drug delivery system. (c) 2009 Wiley Periodicals, Inc.

  16. Synthesis of polypyrrole nanowires with positive effect on MC3T3-E1 cell functions through electrical stimulation.

    Science.gov (United States)

    He, Yuan; Wang, Shihui; Mu, Jing; Dai, Lingfeng; Zhang, Zhong; Sun, Yanan; Shi, Wei; Ge, Dongtao

    2017-02-01

    Conducting polymer polypyrrole (PPy) possesses good biocompatibility and conductivity and has been used as functional coatings in bone tissue regeneration. In this study, a cholic acid doped PPy nanowires (PPy NWs) coating was electrochemically polymerized on the surface of titanium (Ti). The porous intertwined PPy NWs coating exhibited excellent electrical conductivity and electrochemical activity, better hydrophilicity and higher surface energy. In vitro cell experiments demonstrated that the PPy NWs coating together with a 10μA substrate-mediate electrical stimulation (ES) was capable to positive regulate the functions of MC3T3-E1 such as cell adhesion, proliferation and differentiation. Further long-term functions of cell tests including alkaline phosphatase (ALP) activity, bone-carboxyglutamic acid-containing protein (BGP) and calcium deposition were all thoroughly increased. These confirmed that the combination of PPy NWs and ES could accelerate MC3T3-E1 cells mature and osteogenesis. Hence, the PPy NWs coating was an electro bioactive coating and may have potential applications in the treatment of bone damage repairing and regeneration with ES. Copyright © 2016. Published by Elsevier B.V.

  17. Hexon-modified recombinant E1-deleted adenoviral vectors as bivalent vaccine carriers for Coxsackievirus A16 and Enterovirus 71.

    Science.gov (United States)

    Zhang, Chao; Yang, Yong; Chi, Yudan; Yin, Jieyun; Yan, Lijun; Ku, Zhiqiang; Liu, Qingwei; Huang, Zhong; Zhou, Dongming

    2015-09-22

    Hand, foot and mouth disease (HFMD) is a major public health concern in Asia; more efficient vaccines against HFMD are urgently required. Adenoviral (Ad) capsids have been used widely for the presentation of foreign antigens to induce specific immune responses in the host. Here, we describe a novel bivalent vaccine for HFMD based on the hexon-modified, E1-deleted chimpanzee adenovirus serotype 68 (AdC68). The novel vaccine candidate was generated by incorporating the neutralising epitope of Coxsackievirus A16 (CA16), PEP71, into hypervariable region 1 (HVR1), and a shortened neutralising epitope of Enterovirus 71 (EV71), sSP70, into HVR2 of the AdC68 hexon. In order to enhance the immunogenicity of EV71, VP1 of EV71 was cloned into the E1-region of the AdC68 vectors. The results demonstrated that these two epitopes were well presented on the virion surface and had high affinity towards specific antibodies, and VP1 of EV71 was also significantly expressed. In pre-clinical mouse models, the hexon-modified AdC68 elicited neutralising antibodies against both CA16 and EV71, which conferred protection to suckling mice against a lethal challenge of CA16 and EV71. In summary, this study demonstrates that the hexon-modified AdC68 may represent a promising bivalent vaccine carrier against EV71 and CA16 and an epitope-display platform for other pathogens.

  18. NEW ORBITS FOR COMETS C/1960 M1 (HUMASON, C/1980 E1 (BOWELL, AND MUSINGS ON EXTRASOLAR COMETS

    Directory of Open Access Journals (Sweden)

    Richard L. Branham Jr.

    2013-01-01

    Full Text Available Se calculan órbitas nuevas para los cometas Humason (C/1960 M1 y Bowell (C/1980 E1. La órbita de Humason se basa en 34 observaciones hechas durante 348 días y para Bowell en 203 observaciones hechas durante ocho años. Integraciones hacia atrás indican que ambos cometas tenían órbitas originalmente muy elípticas, que fueron cambiadas a hipérbolas por la adición de energía desde el Sistema Solar. Puesto que sus distancias del perihelio son mayores de 3 AU, la posibilidad de fuerzas no gravitatorias es remota. Para el cometa Secchi (C/1853 E1, sin embargo, la órbita es hiperbólica a una distancia de más de 100000 AU del Sol y sin evidencia alguna de fuerzas no gravitatorias. Si la órbita permanece hiperbólica a esa distancia, quizás su origen sea fuera de la nube de Oort.

  19. The role of FIS in the Rcd checkpoint and stable maintenance of plasmid ColE1.

    Science.gov (United States)

    Blaby, I K; Summers, D K

    2009-08-01

    Escherichia coli plasmid ColE1 lacks active partitioning, and copies are distributed randomly to daughter cells at division. The plasmid is maintained stably in the bacterial population as long as its copy number remains high. The accumulation of plasmid dimers and higher multimers depresses copy number, and is an important cause of multicopy plasmid instability. ColE1 dimers are restored to the monomeric state by site-specific recombination, which requires the host-encoded proteins XerCD, ArgR and PepA acting at the plasmid cer site. In addition, a 70 nt RNA expressed from the cer site of plasmid dimers delays the division of dimer-containing cells. Here, we report that the global regulator FIS binds to cer in a sequence-specific manner, close to the Rcd promoter (P(cer)). FIS is not required for plasmid dimer resolution, but is essential for repression of P(cer) in plasmid monomers. Repression also requires the XerCD recombinase, but not ArgR or PepA. We propose a model for monomer-dimer control of P(cer) in which the promoter is repressed in plasmid monomers by the concerted action of FIS and XerCD. Rcd transcription is triggered in plasmid dimers by the lifting of XerCD-mediated repression in the synaptic complex.

  20. Effect of Water Miscible Organic Solvents on p-Nitrophenol Hydroxylase (CYP2E1) Activity in Rat Liver Microsomes.

    Science.gov (United States)

    Patil, Pranali G; Kamble, S H; Shah, T S; Iyer, K R

    2015-01-01

    Organic solvents used for solubilization of the substrates/NCEs are known to affect the activity of cytochrome P450 enzymes. Further, this effect varies with the solvents used, the substrates and CYP450 isoforms in question. In the present study, we have investigated the effect of ten commonly used water miscible organic solvents (methanol, ethanol, 1-propanol, 2-propanol, acetonitrile, acetone, dimethyl sulphoxide, N,N-dimethyl formamide, dioxane and polyethylene glycol 400) on p-nitrophenol hydroxylase activity at 0, 0.1, 0.25, 0.5, 0.75 and 1% v/v concentration in rat liver microsomes. All the solvents studied showed concentration dependent inhibition of the p-nitrophenol hydroxylase activity except acetonitrile which showed activation of the activity at concentration range studied. Out of ten solvents studied, dioxane was found to be the most inhibitory solvent (inhibition >90% at 0.25% v/v concentration). Overall, solvents like dimethyl sulphoxide, dimethyl formamide and dioxane appeared to be unsuitable for characterizing p-nitrophenol hydroxylase (CYP2E1-mediated) reactions due to a high degree of inhibition. On the other hand, methanol and acetonitrile at concentrations <0.5% v/v appeared to be appropriate solvents for substrate solubilization while evaluating CYP2E1-mediated catalysis. The results of this study imply that caution should be exercised while choosing solvents for dissolution of substrate during enzyme studies in liver microsomes.

  1. The global image of the Moon obtained by the Chang’E-1:Data processing and lunar cartography

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The global lunar image of the first phase of Chinese Lunar Exploration Program is the first image that covered all over the surface of the Moon. It will serve as a critical foundation for succeeding exploration and scientific research. In this paper, the acquisition, characteristics, and data quality of Chang’E-1 CCD image data are described in detail. Also described are the methodology and procedure of data processing. According to rule of planetary cartography, the image data have been processed, geometrically corrected, and then mosaicked and merged in a scale of 1:2.5 million. The results of data processing and charting show that the image data of Chang’E-1 CCD and their geometric precision meet the demand of charting a map in the scale of 1:2.5 million. The relative geometric positioning precision of the global image is better than 240 m, and the absolute geometric positioning precision is slightly better than that of the ULCN2005 and Clementine lunar basemap (V2.0). The plane positioning precision is approximately 100-1500 m. This global image proves to be the best global image of the Moon so far in terms of space coverage, image quality, and positioning precision.

  2. Inhibition of adenovirus replication by the E1A antisense transcript initiated from hsp70 and VA-1 promoters.

    Science.gov (United States)

    Miroshnichenko, O I; Borisenko, A S; Ponomareva, T I; Tikhonenko, T I

    1990-03-01

    The E1A region of the adenoviral genome, important for initiation of virus infection and activation of other viral genes, was chosen as a target for engineering antisense RNA (asRNA) to inhibit adenovirus 5 (Ad5) replication in COS-1 cell culture in vitro. The hsp70 promoter, taken from the appropriate heat-shock-protein gene of Drosophila melanogaster, and the VA-1 RNA promoter, derived from the Ad5 gene coding for low-molecular-mass VA-1 RNA and recognized by RNA polymerase III were used as regulatory elements of transcription. The two types of recombinant constructs contained E1A fragments of 710 bp (hsp70 constructs) or 380 or 740 bp (VA-1 RNA constructs) in reverse orientation relative to the promoter position, as well as a transcription termination signal, the SV40 ori, and the gene controlling Geneticin (antibiotic G418) resistance (G418R). After selection of transfected COS-1 cells in the presence of G418, a number of stable G418R cell lines were raised which expressed engineered asRNAs. Plating of Ad5 suspensions of known titre on monolayers of transfected COS-1 cells clearly showed strong inhibition of adenovirus replication by asRNAs: 75% with the hsp70 promoter and 90% with the VA-1 RNA promoter.

  3. Preliminary Scientific Results of Chang'E-1 Lunar Orbiter: Based on Payloads Detection Data in the First Phase

    Institute of Scientific and Technical Information of China (English)

    OUYANG Ziyuan; YANG Jianfeng; ZHANG Wenxi; WANG Jianyu; MOU Lingli; CHANG Jin; ZHANG Liyan; WANG Huanyu; LI Yongquan; ZHANG Xiaohui; ZHENG Yongchun; JIANG Jingshan; WANG Shijin; BIAN Wei; LI Chunlai; SUN Huixian; ZOU Yongliao; LIU Jianzhong; LIU Jianjun; ZHAO Baochang; REN Xin

    2008-01-01

    Chang'E-1 lunar Orbiter was launched by Long March 3A rocket from Xichang Satellite Launch Center at 18:05 BT (Beijing Time) Oct. 24, 2007. It is the first step of its ambitious three-stage moon program, a new milestone in the Chinese space exploration history. The primary science objectives of Chang'E-1 lunar orbiter are to obtain three-Dimension (3D) stereo images of the lunar surface, to analyze the distribution and abundance of elements on the surface, to investigate the thickness of lunar soil, evaluate helium-3 resources and other characteristics, and to detect the space environment around the moon. To achieve the above four mission objectives, eight sets of scientific instruments are chosen as the payloads of the lunar orbiter, including a CCD stereo camera (CCD), a Sagnac-based interferometer spectrometer (IIM), a Laser Altimeter (LAM), a Microwave Radiometer (MRM), a Gamma-Ray Spectrometer (GRS), an X-ray spectrometer (XRS), a High-Energy Particle Detector (HPD), and two Solar Wind Ion Detectors (SWID). The detected data of the payloads show that all payloads work well. This paper introduces the status of payloads in the first phase and preliminary scientific results.

  4. A MUB E2 structure reveals E1 selectivity between cognate ubiquitin E2s in eukaryotes

    Science.gov (United States)

    Lu, Xiaolong; Malley, Konstantin R.; Brenner, Caitlin C.; Koroleva, Olga; Korolev, Sergey; Downes, Brian P.

    2016-08-01

    Ubiquitin (Ub) is a protein modifier that controls processes ranging from protein degradation to endocytosis, but early-acting regulators of the three-enzyme ubiquitylation cascade are unknown. Here we report that the prenylated membrane-anchored ubiquitin-fold protein (MUB) is an early-acting regulator of subfamily-specific E2 activation. An AtMUB3:AtUBC8 co-crystal structure defines how MUBs inhibit E2~Ub formation using a combination of E2 backside binding and a MUB-unique lap-bar loop to block E1 access. Since MUBs tether Arabidopsis group VI E2 enzymes (related to HsUbe2D and ScUbc4/5) to the plasma membrane, and inhibit E2 activation at physiological concentrations, they should function as potent plasma membrane localized regulators of Ub chain synthesis in eukaryotes. Our findings define a biochemical function for MUB, a family of highly conserved Ub-fold proteins, and provide an example of selective activation between cognate Ub E2s, previously thought to be constitutively activated by E1s.

  5. Effect of water miscible organic solvents on p-nitrophenol hydroxylase (CYP2E1 activity in rat liver microsomes

    Directory of Open Access Journals (Sweden)

    Pranali G Patil

    2015-01-01

    Full Text Available Organic solvents used for solubilization of the substrates/NCEs are known to affect the activity of cytochrome P450 enzymes. Further, this effect varies with the solvents used, the substrates and CYP450 isoforms in question. In the present study, we have investigated the effect of ten commonly used water miscible organic solvents (methanol, ethanol, 1-propanol, 2-propanol, acetonitrile, acetone, dimethyl sulphoxide, N,N-dimethyl formamide, dioxane and polyethylene glycol 400 on p-nitrophenol hydroxylase activity at 0, 0.1, 0.25, 0.5, 0.75 and 1% v/v concentration in rat liver microsomes. All the solvents studied showed concentration dependent inhibition of the p-nitrophenol hydroxylase activity except acetonitrile which showed activation of the activity at concentration range studied. Out of ten solvents studied, dioxane was found to be the most inhibitory solvent (inhibition >90% at 0.25% v/v concentration. Overall, solvents like dimethyl sulphoxide, dimethyl formamide and dioxane appeared to be unsuitable for characterizing p-nitrophenol hydroxylase (CYP2E1-mediated reactions due to a high degree of inhibition. On the other hand, methanol and acetonitrile at concentrations <0.5% v/v appeared to be appropriate solvents for substrate solubilization while evaluating CYP2E1-mediated catalysis. The results of this study imply that caution should be exercised while choosing solvents for dissolution of substrate during enzyme studies in liver microsomes.

  6. Gel Microstructure Regulates Proliferation and Differentiation of MC3T3-E1 Cells Encapsulated in Alginate Beads

    Science.gov (United States)

    Lee, Baek-Hee; Li, Bing; Guelcher, Scott A.

    2012-01-01

    For cell transplantation into damaged tissues, viable cells must be delivered to the defect site in a suitable carrier. However, the hypoxic and nutrient-limited environment in the carrier can induce massive cell death. The aims of this study were to increase the viability and regulate the behavior of osteoprogenitor cells encapsulated in alginate hydrogels through control of the gel microstructure. Cell survivability in alginate beads was improved through the use of α-MEM as the solvent for alginic acid sodium salt and CaCl2 solutions, which supplied additional nutrients for the cells compared to water or buffer. The mesh size and shear modulus of the hydrogel were hypothesized to regulate proliferation and differentiation of osteoprogenitor cells. MC3T3-E1 cells demonstrated enhanced osteoblast differentiation when encapsulated in high-density alginate with smaller mesh size and more rigid mechanical properties, as confirmed by increased alkaline phosphatase activity and osteocalcin secretion. However, MC3T3-E1 cells encapsulated in low-density alginate beads with a larger mesh size and more compliant mechanical properties exhibited increased proliferation. These results demonstrate that the microstructure of alginate hydrogels can regulate the behavior of osteoprogenitor cells, thus suggesting that the tuning the properties of the gel may be a useful approach for enhancing new bone formation. PMID:22306825

  7. Reactive oxygen species regulatory mechanisms associated with rapid response of MC3T3-E1 cells for vibration stress

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Ling; Gan, Xueqi; Zhu, Zhuoli; Yang, Yang; He, Yuting; Yu, Haiyang, E-mail: yhyang6812@scu.edu.cn

    2016-02-12

    Although many previous studies have shown that refractory period-dependent memory effect of vibration stress is anabolic for skeletal homeostasis, little is known about the rapid response of osteoblasts simply derived from vibration itself. In view of the potential role of reactive oxygen species (ROS) in mediating differentiated activity of osteoblasts, whether and how ROS regulates the rapid effect of vibration deserve to be demonstrated. Our findings indicated that MC3T3-E1 cells underwent decreased gene expression of Runx2, Col-I and ALP and impaired ALP activity accompanied by increased mitochondrial fission immediately after vibration loading. Moreover, we also revealed the involvement of ERK-Drp1 signal transduction in ROS regulatory mechanisms responsible for the rapid effect of vibration stress. - Highlights: • ROS contributed to the rapid response of MC3T3-E1 cells for vibration stress. • Imbalance of mitochondrial dynamics were linked to the LMHFV-derived rapid response. • The role of ERK-Drp1 signal pathway in the LMHFV-derived osteoblast rapid response.

  8. E1B蛋白缺陷型溶瘤腺病毒对乳腺癌干细胞的作用%Role of E1B protein deficient oncolytic virus in breast cancer stem cells

    Institute of Scientific and Technical Information of China (English)

    马越; 张凤春; 徐迎春; 黄明主; 林叔成; 张雁云

    2008-01-01

    目的 研究E1B蛋白缺陷型溶瘤腺病毒(E1B-- OLV)对乳腺癌干细胞的作用. 方法 E1B- OLV感染人乳腺癌细胞系MCF-7细胞(实验组),常规培养MCF-7细胞作为对照组.两组细胞分别行流式细胞仪(FCM)分析CD44+CD24-细胞的表型比例.两组细胞同时行微球体培养,观察微球体形成的大小及数量,计算微球体形成率(MFE),用FCM分析微球体CD44+CD24-细胞的比例. 结果 实验组MCF-7细胞的CD24-、CD44+和CD44+CD24-比例分别为43.90%、63.26%和22.19%,对照组为6.74%、88.30%和2.30%.实验组中微球体成球时间早于对照组,球体体积大于对照组,MFE高于对照组(1.26%和0.9%);实验组和对照组微球体中CD44+CD24-细胞的比例分别为38.08%和23.35%. 结论 ElB-- OLV在短期内杀死MCF-7细胞,但主要是乳腺癌分化细胞,可能促进了乳腺癌干细胞的生长,加快了其自我更新和分化速率.

  9. Structure, dynamics, and interaction of Mycobacterium tuberculosis (Mtb DprE1 and DprE2 examined by molecular modeling, simulation, and electrostatic studies.

    Directory of Open Access Journals (Sweden)

    Isha Bhutani

    Full Text Available The enzymes decaprenylphosphoryl-β-D-ribose oxidase (DprE1 and decaprenylphosphoryl-β-D-ribose-2-epimerase (DprE2 catalyze epimerization of decaprenylphosporyl ribose (DPR todecaprenylphosporyl arabinose (DPA and are critical for the survival of Mtb. Crystal structures of DprE1 so far reported display significant disordered regions and no structural information is known for DprE2. We used homology modeling, protein threading, molecular docking and dynamics studies to investigate the structural and dynamic features of Mtb DprE1 and DprE2 and DprE1-DprE2 complex. A three-dimensional model for DprE2 was generated using the threading approach coupled with ab initio modeling. A 50 ns simulation of DprE1 and DprE2 revealed the overall stability of the structures. Principal Component Analysis (PCA demonstrated the convergence of sampling in both DprE1 and DprE2. In DprE1, residues in the 269-330 area showed considerable fluctuation in agreement with the regions of disorder observed in the reported crystal structures. In DprE2, large fluctuations were detected in residues 95-113, 146-157, and 197-226. The study combined docking and MD simulation studies to map and characterize the key residues involved in DprE1-DprE2 interaction. A 60 ns MD simulation for DprE1-DprE2 complex was also performed. Analysis of data revealed that the docked complex is stabilized by H-bonding, hydrophobic and ionic interactions. The key residues of DprE1 involved in DprE1-DprE2 interactions belong to the disordered region. We also examined the docked complex of DprE1-BTZ043 to investigate the binding pocket of DprE1 and its interactions with the inhibitor BTZ043. In summary, we hypothesize that DprE1-DprE2 interaction is crucial for the synthesis of DPA and DprE1-DprE2 complex may be a new therapeutic target amenable to pharmacological validation. The findings have important implications in tuberculosis (TB drug discovery and will facilitate drug development efforts against

  10. Interaction of CtBP with adenovirus E1A suppresses immortalization of primary epithelial cells and enhances virus replication during productive infection

    Energy Technology Data Exchange (ETDEWEB)

    Subramanian, T.; Zhao, Ling-jun; Chinnadurai, G., E-mail: chinnag@slu.edu

    2013-09-01

    Adenovirus E1A induces cell proliferation, oncogenic transformation and promotes viral replication through interaction with p300/CBP, TRRAP/p400 multi-protein complex and the retinoblastoma (pRb) family proteins through distinct domains in the E1A N-terminal region. The C-terminal region of E1A suppresses E1A/Ras co-transformation and interacts with FOXK1/K2, DYRK1A/1B/HAN11 and CtBP1/2 (CtBP) protein complexes. To specifically dissect the role of CtBP interaction with E1A, we engineered a mutation (DL→AS) within the CtBP-binding motif, PLDLS, and investigated the effect of the mutation on immortalization and Ras cooperative transformation of primary cells and viral replication. Our results suggest that CtBP–E1A interaction suppresses immortalization and Ras co-operative transformation of primary rodent epithelial cells without significantly influencing the tumorigenic activities of transformed cells in immunodeficient and immunocompetent animals. During productive infection, CtBP–E1A interaction enhances viral replication in human cells. Between the two CtBP family proteins, CtBP2 appears to restrict viral replication more than CtBP1 in human cells. - Highlights: • Adenovirus E1A C-terminal region suppresses E1A/Ras co-transformation. • This E1A region binds with FOXK, DYRK1/HAN11 and CtBP cellular protein complexes. • We found that E1A–CtBP interaction suppresses immortalization and transformation. • The interaction enhances viral replication in human cells.

  11. 基于H.264的E1标准下的视频传输技术改进研究%Improvement Research of Video Transmission Technology Based on H.264 under E1 Standard

    Institute of Scientific and Technical Information of China (English)

    徐宁; 孟凡荣; 孙洋

    2012-01-01

    目前,无线视频传输大多在E1标准下进行,但由于采用了较早的H.261编解码标准,一直存在误码率较高,图像较模糊等问题,影响了指挥机关对现场态势的判断。文章基于先进的H.264编解码标准,采用以TMS320DM6467T高清视频双核媒体处理器作为CPU的SEED-VPM6467独立开发平台,在卫星通信系统中,对无线视频传输技术进行了改进性研究,由过去的CIF(352×288)格式,提升到D1(4CIF)视频格式的传输,对无线视频传输系统,提供了技术依据。%By now,wireless video transmission carried out under E1 standard,but because of the earlier H.263 code and encode standard,there has always been some problems,such as high bit-error rates,pattern blurring,which have affeced the judgement by the military commands according to the situation.This paper puts forward an improvement research of video transmission technology based on advanced H.264 standard,using SEED-VPM6467 as platform,under satelite communications,which could increase the video format from CIF to D1(4CIF).This research provides technical support for wireless video transmission systerm.

  12. 前列腺素E1对移植肝的保护作用%Experiment study on the relationship between the protective effect of prostaglandin E1 and the lysosome in orthotopic liver transplantation in rats

    Institute of Scientific and Technical Information of China (English)

    阚彤; 杨甲梅; 严以群; 陈汉; 吴孟超

    2001-01-01

    Objective To study the protective effect of prostaglandin E1 (PGE1) on the liver in orthotopic liver transplantation rats. Methods PGE1 was infused intravenously during orthotopic liver transplantation in recipient rats. Sham-operated and normal saline groups served as control groups. One-week survival rate and ultrastructure pathological changes in liver were observed. Results The survival rate in the PGE1-treated group was obviously increased as compared with that in the control group. Electron microscopic examination revealed a dramatic elevation of hepatocellular lysosome number in the control groups, and bleb and rupture of lysosome membrane were also observed in the PGE1-trated group. Conclusions PGE1 treatment during recipient operation could increase one-week survival rate, which might be related with its stabilizing effect on lysosome membrane.%目的探讨术中应用前列腺素E1(PGE1)对移植肝脏的保护作用。方法大鼠原位肝移植术中经颈内静脉灌注PGE1,设盐水和空白对照,观察术后1周存活率和肝脏超微结构变化。结果 PGE1治疗组术后1周存活率较对照组明显提高,电镜证实PGE1治疗组肝细胞内溶酶体膜无鼓泡和破溃现象。结论术中应用PGE1能显著提高大鼠肝移植后的1周存活率,其机制可能与PGE1稳定溶酶体膜的作用有关。

  13. Antibody to E1 peptide of hepatitis C virus genotype 4 inhibits virus binding and entry to HepG2 cells in vitro

    OpenAIRE

    2006-01-01

    AIM: To analyze the neutralizing activity of antibodies against E1 region of hepatitis C virus (HCV). Specific polyclonal antibody was raised via immunization of New Zealand rabbits with a synthetic peptide that had been derived from the E1 region of HCV and was shown to be highly conserved among HCV published genotypes.

  14. 26 CFR 1.417(e)-1 - Restrictions and valuations of distributions from plans subject to sections 401(a)(11) and 417.

    Science.gov (United States)

    2010-04-01

    ... from plans subject to sections 401(a)(11) and 417. 1.417(e)-1 Section 1.417(e)-1 Internal Revenue... from plans subject to sections 401(a)(11) and 417. (a) Scope—(1) In general. A plan does not satisfy... the account balance, a defined contribution plan is not subject to the requirements of this...

  15. The design, synthesis and biological evaluation of novel thiamin diphosphate analog inhibitors against the pyruvate dehydrogenase multienzyme complex E1 from Escherichia coli.

    Science.gov (United States)

    Feng, Lingling; He, Junbo; He, Haifeng; Zhao, Lulu; Deng, Lingfu; Zhang, Li; Zhang, Lin; Ren, Yanliang; Wan, Jian; He, Hongwu

    2014-11-28

    Pyruvate dehydrogenase multienzyme complex E1 (PDHc E1) is a potential target enzyme when looking for inhibitors to combat microbial disease. In this study, we designed and synthesized a series of novel thiamin diphosphate (ThDP) analogs with triazole ring and oxime ether moieties as potential inhibitors of PDHc E1. Their inhibitory activities against PDHc E1 were examined both in vitro and in vivo. Most of the tested compounds exhibited moderate inhibitory activities against PDHc E1 (IC50 = 6.1-75.5 μM). The potent inhibitors 4g, 4h and 4j, had strong inhibitory activities with IC50 values of 6.7, 6.9 and 6.1 μM against PDHc E1 in vitro and with inhibition rates of 35%, 50% and 33% at 100 μg mL(-1) against Gibberella zeae in vivo, respectively. The binding mode of 4j to PDHc E1 was analyzed by a molecular docking method. Furthermore, the possible interactions of the important residues of PDHc E1 with compound 4j were examined by site-directed mutagenesis, enzymatic assays and spectral fluorescence studies. The theoretical and experimental results are in good agreement and suggest that compound 4j could be used as a lead compound for further optimization, and may have potential as a new microbicide.

  16. The olfactory receptor OR51E1 is present along the gastrointestinal tract of pigs and is modulated by intestinal microbiota

    NARCIS (Netherlands)

    Priori, D.; Clavenzani, P.; Jansman, A.J.M.; Lalles, J.P.; Trivisil, P.; Bosi, P.

    2015-01-01

    The relevance of the butyrate-sensing olfactory receptor OR51E1 for gastrointestinal (GIT) functioning has not been considered so far. We investigated in young pigs the distribution of OR51E1 along the GIT, its relation with some endocrine markers, its variation with age and after interventions affe

  17. 29 CFR 102.83 - Petition for referendum under section 9(e)(1) of the Act; who may file; where to file; withdrawal.

    Science.gov (United States)

    2010-07-01

    ... 29 Labor 2 2010-07-01 2010-07-01 false Petition for referendum under section 9(e)(1) of the Act... NATIONAL LABOR RELATIONS BOARD RULES AND REGULATIONS, SERIES 8 Procedure for Referendum Under Section 9(e) of the Act § 102.83 Petition for referendum under section 9(e)(1) of the Act; who may file; where to...

  18. Retina restored and brain abnormalities ameliorated by single-copy knock-in of human NR2E1 in null mice.

    Science.gov (United States)

    Schmouth, J-F; Banks, K G; Mathelier, A; Gregory-Evans, C Y; Castellarin, M; Holt, R A; Gregory-Evans, K; Wasserman, W W; Simpson, E M

    2012-04-01

    Nr2e1 encodes a stem cell fate determinant of the mouse forebrain and retina. Abnormal regulation of this gene results in retinal, brain, and behavioral abnormalities in mice. However, little is known about the functionality of human NR2E1. We investigated this functionality using a novel knock-in humanized-mouse strain carrying a single-copy bacterial artificial chromosome (BAC). We also documented, for the first time, the expression pattern of the human BAC, using an NR2E1-lacZ reporter strain. Unexpectedly, cerebrum and olfactory bulb hypoplasia, hallmarks of the Nr2e1-null phenotype, were not fully corrected in animals harboring one functional copy of human NR2E1. These results correlated with an absence of NR2E1-lacZ reporter expression in the dorsal pallium of embryos and proliferative cells of adult brains. Surprisingly, retinal histology and electroretinograms demonstrated complete correction of the retina-null phenotype. These results correlated with appropriate expression of the NR2E1-lacZ reporter in developing and adult retina. We conclude that the human BAC contained all the elements allowing correction of the mouse-null phenotype in the retina, while missing key regulatory regions important for proper spatiotemporal brain expression. This is the first time a separation of regulatory mechanisms governing NR2E1 has been demonstrated. Furthermore, candidate genomic regions controlling expression in proliferating cells during neurogenesis were identified.

  19. E2F activates late-G1 events but cannot replace E1A in inducing S phase in terminally differentiated skeletal muscle cells

    DEFF Research Database (Denmark)

    Pajalunga, D; Tognozzi, D; Tiainen, M

    1999-01-01

    We have previously shown that the adenovirus E1A oncogene can reactivate the cell cycle in terminally differentiated cells. Current models imply that much or all of this E1A activity is mediated by the release of the E2F transcription factors from pocket-protein control. In contrast, we show here...

  20. Deacylation of the transmembrane domains of Sindbis virus envelope glycoproteins E1 and E2 does not affect low-pH-induced viral membrane fusion activity

    NARCIS (Netherlands)

    Smit, JM; Bittman, R; Wilschut, J

    2001-01-01

    The envelope glycoproteins E1 and E2 of Sindbis virus are palmitoylated at cysteine residues within their transmembrane domains (E1 at position 430, and E2 at positions 388 and 390), Here, we investigated the in vitro membrane fusion activity of Sindbis virus variants (derived from the Tote 1101 inf

  1. Specifically Binding of L-ficolin to N-glycans of HCV Envelope Glycoproteins E1 and E2 Leads to Complement Activation

    Institute of Scientific and Technical Information of China (English)

    Jun Liu; Mohammed A.M.Ali; Yinghua Shi; Yinglan Zhao; Fenglin Luo; Jin Yu; Tian Xiang; Jie Tang; Dongqing Li; Quan Hu; Wenzhe Ho; Xiaolian Zhang

    2009-01-01

    L-ficolin and HCV E1 and E2 glycoproteins was attributed to the N-glycans of E1 and E2.These findings provide new insights into the biological functions of L-ficolin in clinically important hepatic viral diseases.

  2. Transgenic plants of Petunia hybrida harboring the CYP2E1 gene efficiently remove benzene and toluene pollutants and improve resistance to formaldehyde

    Directory of Open Access Journals (Sweden)

    Daoxiang Zhang

    2011-01-01

    Full Text Available The CYP2E1 protein belongs to the P450 enzymes family and plays an important role in the metabolism of small molecular and organic pollutants. In this study we generated CYP2E1 transgenic plants of Petunia using Agrobacterium rhizogenes K599. PCR analysis confirmed that the regenerated plants contained the CYP2E1 transgene and the rolB gene of the Ri plasmid. Southern blotting revealed the presence of multiple copies of CYP2E1 in the genome of transgenic plants. Fluorescent quantitative PCR revealed exogenous CYP2E1 gene expression in CYP2E1 transgenic plants at various levels, whereas no like expression was detected in either GUS transgenic plants or wild-types. The absorption of benzene and toluene by transgenic plants was analyzed through quantitative gas chromatography. Transgenic plants with high CYP2E1 expression showed a significant increase in absorption capacity of environmental benzene and toluene, compared to control GUS transgenic and wild type plants. Furthermore, these plants also presented obvious improved resistance to formaldehyde. This study, besides being the first to reveal that the CYP2E1 gene enhances plant resistance to formaldehyde, also furnishes a new method for reducing pollutants, such as benzene, toluene and formaldehyde, by using transgenic flowering horticultural plants.

  3. Reinvestigation of the high spin states in ^(161)Er and enhanced E1 transitions in the N=93 isotones

    Institute of Scientific and Technical Information of China (English)

    陈亮; 周厚兵; 丁兵; 王海霞; 郑勇; 竺礼华; 吴晓光; 周小红; 张玉虎; 柳敏良; 王世陶; 方永得; 滑伟; 强赟华; 李广顺

    2011-01-01

    High-spin states in 161Er have been studied experimentally using the 150Nd(16O, 5n) reaction at a beam energy of 86 MeV. The relatively enhanced E1 transitions between the 5/2+[642] and 3/2-[521] bands are observed in 161Er, and the B(E1) values are extra

  4. Preparation of E-1,3-diaminoethenyl functional groups by the reaction of enol tosylate of alpha-formylglycine with primary and secondary amines

    Directory of Open Access Journals (Sweden)

    Sitaram Bhavaraju

    2016-03-01

    Full Text Available The E-1,3-diaminoethenyl functional group is a potentially useful synthon. A number of examples of E-1,3-diaminoethenyl functional groups were prepared in good yield starting from an E-enol tosylate of a serine based diketopiperazine and 1°- or 2° amine nucleophiles. The reaction proceeds via a stereoselective nucleophilic substitution pathway.

  5. Hyperactivity, startle reactivity and cell-proliferation deficits are resistant to chronic lithium treatment in adult Nr2e1frc/frc mice

    Science.gov (United States)

    Wong, Bibiana K.Y.; Hossain, Sazzad M.; Trinh, Eric; Ottmann, Glen A.; Budaghzadeh, Saeed; Zheng, Qing Y.; Simpson, Elizabeth M.

    2012-01-01

    The NR2E1 region on Chromosome 6q21-22 has been repeatedly linked to bipolar disorder (BP) and NR2E1 has been associated with BP, and more specifically bipolar I disorder (BPI). In addition, patient sequencing has revealed an enrichment of rare candidate-regulatory variants. Interestingly, mice carrying either spontaneous (Nr2e1frc) or targeted (Tlx−) deletions of Nr2e1 (here collectively known as Nr2e1-null) show similar neurological and behavioral anomalies, including: hypoplasia of the cerebrum, reduced neural stem cell proliferation, extreme aggression, and deficits in fear conditioning; traits that have been observed in some patients with BP. Thus, NR2E1 is a positional and functional candidate for a role in BP. However, no Nr2e1-null mice have been fully evaluated for behaviors used to model BP in rodents or pharmacological responses to drugs effective in treating BP symptoms. In this study we examine Nr2e1frc/frc mice, homozygous for the spontaneous deletion, for abnormalities in activity, learning and information processing, and cell proliferation; phenotypes that are either affected in patients with BP or commonly assessed in rodent models of BP. The effect of lithium, a drug used to treat BP, was also evaluated for its ability to attenuate Nr2e1frc/frc behavioral and neural stem cell proliferation phenotypes. We show for the first time that Nr2e1-null mice exhibit extreme hyperactivity in the open field as early as postnatal day 18 and in the home cage, deficits in open-field habituation and passive avoidance, and, surprisingly, an absence of acoustic startle. We observed a reduction in neural stem/progenitor cell proliferation in Nr2e1frc/frc mice, similar to that seen in other Nr2e1-null strains. These behavioral and cell-proliferation phenotypes were resistant to chronic-adult-lithium treatment. Thus, Nr2e1frc/frc mice exhibit behavioral traits used to model BP in rodents, but our results do not support Nr2e1frc/frc mice as pharmacological models for

  6. Structure of the retinoblastoma protein bound to adenovirus E1A reveals the molecular basis for viral oncoprotein inactivation of a tumor suppressor

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Xin; Marmorstein, Ronen (UPENN)

    2008-04-02

    The adenovirus (Ad) E1A (Ad-E1A) oncoprotein mediates cell transformation, in part, by displacing E2F transcription factors from the retinoblastoma protein (pRb) tumor suppressor. In this study we determined the crystal structure of the pRb pocket domain in complex with conserved region 1 (CR1) of Ad5-E1A. The structure and accompanying biochemical studies reveal that E1A-CR1 binds at the interface of the A and B cyclin folds of the pRb pocket domain, and that both E1A-CR1 and the E2F transactivation domain use similar conserved nonpolar residues to engage overlapping sites on pRb, implicating a novel molecular mechanism for pRb inactivation by a viral oncoprotein.

  7. Transcriptional repression of hDaxx enhanced by adenovirus 12 E1B 55-kDa oncoprotein interacting with hDaxx

    Institute of Scientific and Technical Information of China (English)

    万艳平; 吴移谋; 朱翠明; 尹卫国; 蔡恒玲; 余敏君

    2004-01-01

    Background Daxx has been identified as a nuclear protein that involves in apoptosis and transcriptional repression. Daxx co-localizes with the promyelocytic leukemia (PML) protein and regulates transcription. Human Daxx (hDaxx) is a protein that functions as a transcriptional regulation through its interaction with some DNA-associated proteins. The aim of this study was to explore the transcriptional regulatory effect of hDaxx interacting with adenovirus (Ad) 12 E1B (Ad12E1B) 55-kDa oncoprotein.Methods The co-localization of hDaxx-Ad12E1B or hDaxx-PML protein in the nucleus was observed under a confocal microscope. Interaction of hDaxx and Ad12E1B was analyzed by yeast two-hybrid assay. Direct binding of hDaxx and Ad12E1B was analyzed using coimmunoprecipitation and Western blot in vivo and in vitro. The activity of a luciferase reporter gene, which was regulated by an hDaxx modulated thymidine kinase (TK) promoter, was detected in an automat luminometer.Results Ad12E1B, which co-localized with hDaxx in the nuclei of G401-CC3 cells, disrupted the co-localization of hDaxx and PML in the PML oncogenic domains (PODs). hDaxx bound directly to Ad12E1B in vivo and in vitro. hDaxx interacted with Ad12E1B along its full length. Ad12E1B enhanced transcriptional repression activity of hDaxx.Conclusion Ad12E1B disrupts the co-localization of hDaxx with PML in PODs and enhances transcriptional repression activity of hDaxx.

  8. Poly-γ-glutamic acid productivity of Bacillus subtilis BsE1 has positive function in motility and biocontrol against Fusarium graminearum.

    Science.gov (United States)

    Wang, Luyao; Wang, Ning; Mi, Dandan; Luo, Yuming; Guo, Jianhua

    2017-07-01

    In this study, we investigate the relationship between γ-PGA productivity and biocontrol capacity of Bacillus subtilis BsE1; one bacterial isolate displayed 62.14% biocontrol efficacy against Fusarium root rot. The γ-PGA yield assay, motility assay, wheat root colonization assay, and biological control assay were analysed in different γ-PGA yield mutants of BsE1. The pgsB (PGA-synthase-CapB gene) deleted mutant of BsE1 reduced γ-PGA yield and exhibited apparent decline of in vitro motile ability. Deletion of pgsB impaired colonizing capacity of BsE1 on wheat root in 30 days, also lowered biocontrol efficacies from 62.08% (wild type BsE1) to 14.22% in greenhouse experiment against Fusarium root rot. The knockout of pgdS and ggt (genes relate to two γ-PGA degrading enzymes) on BsE1, leads to a considerable improvement in polymer yield and biocontrol efficacy, which attains higher level compared with wild type BsE1. Compared with ΔpgsB mutant, defense genes related to reactive oxygen species (ROS) and phytoalexin expressed changes by notable levels on wheat roots treated with BsE1, demonstrating the functional role γ-PGA plays in biocontrol against Fusarium root rot. γ-PGA is not only important to the motile and plant root colonization ability of BsE1, but also essential to the biological control performed by BsE1 against Fusarium root rot. Our goal in this study is to reveals a new perspective of BCAs screening on bacterial isolates, without good performance during pre-assays of antagonism ability.

  9. Metabolism of methyl tert-butyl ether and other gasoline ethers in mouse liver microsomes lacking cytochrome P450 2E1.

    Science.gov (United States)

    Hong, J Y; Wang, Y Y; Bondoc, F Y; Yang, C S; Gonzalez, F J; Pan, Z; Cokonis, C D; Hu, W Y; Bao, Z

    1999-03-08

    To reduce the production of pollutants in motor vehicle exhaust, methyl tert-butyl ether (MTBE) and other ethers such as ethyl tert-butyl ether (ETBE) and tert-amyl methyl ether (TAME) are added to gasoline as oxygenates for more complete combustion. Metabolism of these gasoline ethers is catalyzed by cytochrome P450 (P450) enzymes. P450 2E1, which metabolizes diethyl ether, was suggested to be an enzyme involved. The present study used 2E1 knock-out mice (2E1-/-) to assess the contribution of 2E1 to the metabolism of MTBE, ETBE and TAME. Liver microsomes prepared from the 2E1 knock-out mice lacked 2E1 activity (assayed as N-nitrosodimethylamine demethylation), but were still active in metabolizing all three gasoline ethers. The levels of ether-metabolizing activity (nmol/min per mg) in the liver microsomes from 7 week old female 2E1 knock-out mice were 0.54+/-0.17 for MTBE, 0.51+/-0.24 for ETBE and 1.14+/-0.25 for TAME at a 1 mM substrate concentration. These activity levels were not significantly different from those of the sex- and age-matched C57BL/6N and 129/Sv mice, which are the parental lineage strains of the 2E1 knock-out mice and are both 2E1+/+. Our results clearly demonstrate that 2E1 plays a negligible role in the metabolism of MTBE, ETBE and TAME in mouse livers.

  10. Isolation and characterization of broadly neutralizing human monoclonal antibodies to the e1 glycoprotein of hepatitis C virus

    DEFF Research Database (Denmark)

    Meunier, Jean-Christophe; Russell, Rodney S.; Goossens, Vera

    2008-01-01

    The relative importance of humoral and cellular immunity in the prevention or clearance of hepatitis C virus (HCV) infection is poorly understood. However, there is considerable evidence that neutralizing antibodies are involved in disease control. Here we describe the detailed analysis of human...... monoclonal antibodies (MAbs) directed against HCV glycoprotein E1, which may have the potential to control HCV infection. We have identified two MAbs that can strongly neutralize HCV-pseudotyped particles (HCVpp) bearing the envelope glycoproteins of genotypes 1a, 1b, 4a, 5a, and 6a and less strongly...... neutralize HCVpp bearing the envelope glycoproteins of genotype 2a. Genotype 3a was not neutralized. The epitopes for both MAbs were mapped to the region encompassing amino acids 313 to 327. In addition, robust neutralization was also observed against cell culture-adapted viruses of genotypes 1a and 2a...

  11. An automated exploration of the isomerization and dissociation pathways of (E)-1,2-dichloroethene cations and anions

    Science.gov (United States)

    Kishimoto, Naoki; Nishi, Yuito

    2017-04-01

    Isomerization and dissociation pathways after the photoionization or electron attachment of (E)-1,2-dichloroethene were calculated with an automated exploration method utilizing a scaled hypersphere search of the anharmonic downward distortion following algorithm at the UB3LYP/6-311G(2d,d,p) level of theory. The potential energies of transition states and dissociation channels were calculated by a composite method ((RO)CBS-QB3) and compared with the breakdown diagrams and electron attachment spectra observed in previous spectroscopic studies. The results of single point calculations with several DFT and post-SCF methods are compared using the root mean square deviations from the (RO)CBS-QB3 energies for six states of anionic dichloroethene.

  12. Effect of injection molded micro-structured polystyrene surfaces on proliferation of MC3T3-E1 cells

    Directory of Open Access Journals (Sweden)

    G. Lucchetta

    2015-04-01

    Full Text Available In this work, osteoinductive micro-pillared polystyrene surfaces were mass-produced for bone replacement applications, by means of the micro injection molding process. Firstly, the molding process parameters were optimized with a two-level, three-factor central composite face-centered plan to increase the quality of polystyrene micro pillars replication and to maximize the pillars height uniformity over the molded part. Secondly, osteoblastic MC3T3-E1 cells adhesion and proliferation on the replicated substrates were assessed as a function of micro topography parameters, such as pillars diameter, aspect ratio and spacing. Cell morphology and proliferation were evaluated through MTS test after 1, 3 and 7 days from seeding. The experimental results showed that cells adhesion and proliferation is more positively promoted on micro-pillared surfaces compared to flat surfaces, but no correlations were observed between cell proliferation and pillar diameter and spacing.

  13. A Field Trial of TCE Phytoremediation by Genetically Modified Poplars Expressing Cytochrome P450 2E1.

    Science.gov (United States)

    Legault, Emily K; James, C Andrew; Stewart, Keith; Muiznieks, Indulis; Doty, Sharon L; Strand, Stuart E

    2017-06-06

    A controlled field study was performed to evaluate the effectiveness of transgenic poplars for phytoremediation. Three hydraulically contained test beds were planted with 12 transgenic poplars, 12 wild type (WT) poplars, or left unplanted, and dosed with equivalent concentrations of trichloroethylene (TCE). Removal of TCE was enhanced in the transgenic tree bed, but not to the extent of the enhanced removal observed in laboratory studies. Total chlorinated ethene removal was 87% in the CYP2E1 bed, 85% in the WT bed, and 34% in the unplanted bed in 2012. Evapotranspiration of TCE from transgenic leaves was reduced by 80% and diffusion of TCE from transgenic stems was reduced by 90% compared to WT. Cis-dichloroethene and vinyl chloride levels were reduced in the transgenic tree bed. Chloride ion accumulated in the planted beds corresponding to the TCE loss, suggesting that contaminant dehalogenation was the primary loss fate.

  14. ZIP1 and zinc inhibits fluoride-induced apoptosis in MC3T3-E1 cells.

    Science.gov (United States)

    Xu, Shihong; Yang, Yongliang; Han, Shumei; Wu, Zonghui

    2014-06-01

    Excess fluoride intake could induce apoptosis in the cells. As an essential micronutrient and cytoprotectant, zinc is involved in many types of apoptosis. Here, we studied the effects of zinc and ZIP1 on fluoride-induced apoptosis in mouse MC3T3-E1 cells and examined the underlying molecular mechanisms. Our study found that fluoride not only inhibited cell proliferation and increased the intracellular reactive oxygen species (ROS) but also induced cell apoptosis. Whereas pretreatment with zinc significantly attenuated fluoride-induced ROS production and partly protected cells against fluoride-induced apoptosis through MAPK/ERK signaling pathway. Our study also found that fluoride upregulated the expression of ZIP1 in a time-dependent manner. Moreover, overexpression of ZIP1 also inhibited fluoride-induced apoptosis by activation of PI3K/Akt pathway. This cytoprotective effect of zinc and ZIP1 may be new factors that affect the physiological activity of fluoride and need study further.

  15. Super-resolution reconstruction and higher-degree function deformation model based matching for Chang’E-1 lunar images

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    This article intends to solve the matching problem of 2C level lunar images by Chang’E-1(CE-1)lunar probe satellite.A line-scanner image matching method is proposed which represents deformation by the quadric function along the camera motion direction and bases on the deformation model for a relief terrain’s imaging on sensors of the satellite borne three-line scanner camera.A precise matching is carried out for the normal view,the frontward view,and the backward view images of the CE-1 by combining the proposed method with the standard correlation method.A super-resolution(SR)reconstruction algorithm based on the wavelet interpolation of non-uniformly sampled data is also adopted to realize SR reconstruction of CE-1 lunar images,which adds the recognizable targets and explores CE-1 lunar images to the full.

  16. Super-resolution reconstruction and higher-degree function deformation model based matching for Chang'E-1 lunar images

    Institute of Scientific and Technical Information of China (English)

    LI LiChun; YU QiFeng; YUAN Yun; SHANG Yang; LU HongWei; SUN XiangYi

    2009-01-01

    This article intends to solve the matching problem of 2C level lunar images by Chang'E-1(CE-1)lunar probe satellite.A line-scanner image matching method is proposed which represents deformation by the quadric function along the camera motion direction and bases on the deformation model for a relief terrain's imaging on sensors of the satellite borne three-line scanner camera.A precise matching is carried out for the normal view,the frontward view,and the backward view images of the CE-1 by combining the proposed method with the standard correlation method.A super-resolution(SR)reconatruction algorithm based on the wavelet interpolation of non-uniformly sampled data is also adopted to realize SR reconstruction of CE-1 lunar images,which adds the recognizable targets and explores CE-1 lunar images to the full.

  17. KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification

    DEFF Research Database (Denmark)

    Bürck, Carolin; Mund, Andreas; Berscheminski, Julia

    2016-01-01

    characterized, but represent a decisive moment in establishing a productive infection. Here, we identify a novel host viral restriction factor, KAP1. This heterochromatin associated transcription factor regulates the dynamic organization of host chromatin structure via its ability to influence epigenetic marks...... epigenetic gene silencing and to promote SUMO modification of E1B-55K by a so far unknown mechanism. IMPORTANCE: Here we describe a novel cellular restriction factor for Human Adenovirus (HAdV) that sheds light on very early modulation processes in viral infection. We reported that chromatin formation...... and cellular SWI/SNF chromatin remodeling play a key role in HAdV transcriptional regulation (1-4). We observed that the cellular chromatin-associated factor, and epigenetic reader SPOC1 represses HAdV infection and gene expression. Here, we illustrate the role of the SPOC1 interacting factor KAP1 during...

  18. Adiponectin and AMP kinase activator stimulate proliferation, differentiation, and mineralization of osteoblastic MC3T3-E1 cells

    Directory of Open Access Journals (Sweden)

    Yamauchi Mika

    2007-11-01

    Full Text Available Abstract Background Adiponectin is a key mediator of the metabolic syndrome that is caused by visceral fat accumulation. Adiponectin and its receptors are known to be expressed in osteoblasts, but their actions with regard to bone metabolism are still unclear. In this study, we investigated the effects of adiponectin on the proliferation, differentiation, and mineralization of osteoblastic MC3T3-E1 cells. Results Adiponectin receptor type 1 (AdipoR1 mRNA was detected in the cells by RT-PCR. The adenosine monophosphate-activated protein kinase (AMP kinase was phosphorylated by both adiponectin and a pharmacological AMP kinase activator, 5-amino-imidazole-4-carboxamide-riboside (AICAR, in the cells. AdipoR1 small interfering RNA (siRNA transfection potently knocked down the receptor mRNA, and the effect of this knockdown persisted for as long as 10 days after the transfection. The transfected cells showed decreased expressions of type I collagen and osteocalcin mRNA, as determined by real-time PCR, and reduced ALP activity and mineralization, as determined by von Kossa and Alizarin red stainings. In contrast, AMP kinase activation by AICAR (0.01–0.5 mM in wild-type MC3T3-E1 cells augmented their proliferation, differentiation, and mineralization. BrdU assay showed that the addition of adiponectin (0.01–1.0 μg/ml also promoted their proliferation. Osterix, but not Runx-2, appeared to be involved in these processes because AdipoR1 siRNA transfection and AICAR treatments suppressed and enhanced osterix mRNA expression, respectively. Conclusion Taken together, this study suggests that adiponectin stimulates the proliferation, differentiation, and mineralization of osteoblasts via the AdipoR1 and AMP kinase signaling pathways in autocrine and/or paracrine fashions.

  19. Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism

    Directory of Open Access Journals (Sweden)

    Węgrzyn Alicja

    2006-11-01

    Full Text Available Abstract Background Although understanding of physiological interactions between plasmid DNA and its host is important for vector design and host optimization in many biotechnological applications, to our knowledge, global studies on plasmid-host interactions have not been performed to date even for well-characterized plasmids. Results Escherichia coli cells, either devoid of plasmid DNA or bearing plasmid pOri1 (with a single ColE1 replication origin or plasmid pOri2 (with double ColE1 replication origins, were cultured in a chemostat. We used a combination of metabolic flux analysis, DNA microarray and enzyme activity analysis methods to explore differences in the metabolism between these strains. We found that the presence of plasmids significantly influenced various metabolic pathways in the host cells, e.g. glycolysis, the tricarboxylic acid (TCA cycle and the pentose phosphate (PP pathway. Expression of rpiA, a gene coding for ribose-5-phosphate isomerase A, was considerably decreased in E. coli carrying a high copy number plasmid relative to E. coli carrying a low copy number plasmid and plasmid-free E. coli. The rpiA gene was cloned into an expression vector to construct plasmid pETrpiA. Following induction of pETrpiA-bearing E. coli, which harbored either pOri1 or pOri2, with isopropyl-β-D-thiogalactopyranoside (IPTG, the copy number of pOri1 and pOri2 was sigificantly higher than that measured in a host devoid of pETrpiA. Conclusion The presence of plasmids can significantly influence some metabolic pathways in the host cell. We believe that the results of detailed metabolic analysis may be useful in optimizing host strains, vectors and cultivation conditions for various biotechnological purposes.

  20. DMSO is a strong inducer of DNA hydroxymethylation in pre-osteoblastic MC3T3-E1 cells

    Science.gov (United States)

    Thaler, Roman; Spitzer, Silvia; Karlic, Heidrun; Klaushofer, Klaus; Varga, Franz

    2012-01-01

    Artificial induction of active DNA demethylation appears to be a possible and useful strategy in molecular biology research and therapy development. Dimethyl sulfoxide (DMSO) was shown to cause phenotypic changes in embryonic stem cells altering the genome-wide DNA methylation profiles. Here we report that DMSO increases global and gene-specific DNA hydroxymethylation levels in pre-osteoblastic MC3T3-E1 cells. After 1 day, DMSO increased the expression of genes involved in DNA hydroxymethylation (TET) and nucleotide excision repair (GADD45) and decreased the expression of genes related to DNA methylation (Dnmt1, Dnmt3b, Hells). Already 12 hours after seeding, before first replication, DMSO increased the expression of the pro-apoptotic gene Fas and of the early osteoblastic factor Dlx5, which proved to be Tet1 dependent. At this time an increase of 5-methyl-cytosine hydroxylation (5-hmC) with a concomitant loss of methyl-cytosines on Fas and Dlx5 promoters as well as an increase in global 5-hmC and loss in global DNA methylation was observed. Time course-staining of nuclei suggested euchromatic localization of DMSO induced 5-hmC. As consequence of induced Fas expression, caspase 3/7 and 8 activities were increased indicating apoptosis. After 5 days, the effect of DMSO on promoter- and global methylation as well as on gene expression of Fas and Dlx5 and on caspases activities was reduced or reversed indicating down-regulation of apoptosis. At this time, up regulation of genes important for matrix synthesis suggests that DMSO via hydroxymethylation of the Fas promoter initially stimulates apoptosis in a subpopulation of the heterogeneous MC3T3-E1 cell line, leaving a cell population of extra-cellular matrix producing osteoblasts.  PMID:22507896

  1. Cytochrome P450 2E1 high activity polymorphism in alcohol abuse and end-organ disease

    Institute of Scientific and Technical Information of China (English)

    Mark T Cartmell; Hans-Ulrich Schulz; Derek A O'Reilly; Bing-Mei Yang; Volker Kielstein; Simon P Dunlop; Walter Halangk; Andrew G Demaine; Andrew N Kingsnorth

    2005-01-01

    AIM: To investigate a possible role for a recently identified polymorphism in the gene of cytochrome P450 2E1, the presence of which is associated with high activity of the enzyme. METHODS: Two hundred and thirty-nine alcohol consumers, ICD 10.1/.2 (ALC), and 208 normal controls were studied. PCR amplification of the CYP2E1 gene region was performed to assess polymorphic variation. Fisher's exact test was used to assess the data.RESULTS: Twelve normal controls (5.8%) possessed the insertion. Five ALC (2.1%) had the insertion; of these 2 of 144 with,alcohol induced chronic pancreatitis, none of 28 with alcoholic liver disease and 3 of 67 without endorgan disease had the polymorphism. A significantly Lower frequency of subjects possessed the insertion than normal controls [P = 0.049 (genotype analysis P = 0.03)]. To further assess, if there was a relationship to alcohol problems per se or end-organ disease, we compared patients with alcohol induced end-organ disease vs alcoholic controls without end-organ disease vs normal controls which again showed a significant difference [P = 0.045 (genotype analysis,P = 0.011)], further sub-group analysis did not identify which group(s) accounted for these differences.CONCLUSION: We have shown the frequencies of this high-activity polymorphism in alcohol related patient groups for the first time. The frequency is significantly less in alcoholics than normal controls, as with high activity polymorphisms of alcohol dehydrogenase. The biological significance, and whether the relevance is solely for alcoholism or is there a relationship to end-organ disease,would benefit from the assessment in the populations with a greater frequency of this polymorphism.

  2. Accurate Registration of the CHANG'E-1 Iim Data Based on Lro Lroc-Wac Mosica Data

    Science.gov (United States)

    Yang, Z.; Kang, Z.

    2017-07-01

    In the detection of the moon, the visible and near-infrared reflectance data of the lunar material are important information sources for lunar chemical substances and mineral inversion. The Interferometer Imaging Spectrometer (IIM) aboard the Chang'E-1 lunar orbiter is the first multispectral imaging spectrometer for Chinese lunar missions. In this paper, we use the mosaic image of global moon acquired by the Wide-angle Camera (WAC) of the Lunar Reconnaissance Orbiter Camera (LROC) to realize the accurate registration of Chang'E-1 IIM hyperspectral images. Due to the lack of GCPs, the emphasis of this work is to find a huge number of homologous points. The method proposed in this paper is to obtain several homologous points by manually matching, and then we utilize those points to calculate the initial homography matrix of LROC-WAC image and IIM image. This matrix is used to predict the area on IIM image where homologous points may be located, and the locations of the homologous points are determined by the orientation correlation in frequency domain. Finally we save the parts of homologous points which satisfied the conversion relationship of initial homography matrix to calculate homography matrix again. We use this iterative way to obtain a more accurate location of the homologous points. In this process, we take into account that the geometric deformations of different regions on IIM image are quite different. Therefore, we added image threshold segmentation based on the initial homography matrix in the experiment, and completed the above work of finding the homologous points on the segmented images. The final realization of registration accuracy of IIM images are in 1-2 pixels (RMSE). This provides a reliable data assurance for the subsequent study of using IIM images to inverse the lunar elements.

  3. ACCURATE REGISTRATION OF THE CHANG’E-1 IIM DATA BASED ON LRO LROC-WAC MOSICA DATA

    Directory of Open Access Journals (Sweden)

    Z. Yang

    2017-07-01

    Full Text Available In the detection of the moon, the visible and near-infrared reflectance data of the lunar material are important information sources for lunar chemical substances and mineral inversion. The Interferometer Imaging Spectrometer (IIM aboard the Chang'E-1 lunar orbiter is the first multispectral imaging spectrometer for Chinese lunar missions. In this paper, we use the mosaic image of global moon acquired by the Wide-angle Camera (WAC of the Lunar Reconnaissance Orbiter Camera (LROC to realize the accurate registration of Chang'E-1 IIM hyperspectral images. Due to the lack of GCPs, the emphasis of this work is to find a huge number of homologous points. The method proposed in this paper is to obtain several homologous points by manually matching, and then we utilize those points to calculate the initial homography matrix of LROC-WAC image and IIM image. This matrix is used to predict the area on IIM image where homologous points may be located, and the locations of the homologous points are determined by the orientation correlation in frequency domain. Finally we save the parts of homologous points which satisfied the conversion relationship of initial homography matrix to calculate homography matrix again. We use this iterative way to obtain a more accurate location of the homologous points. In this process, we take into account that the geometric deformations of different regions on IIM image are quite different. Therefore, we added image threshold segmentation based on the initial homography matrix in the experiment, and completed the above work of finding the homologous points on the segmented images. The final realization of registration accuracy of IIM images are in 1–2 pixels (RMSE. This provides a reliable data assurance for the subsequent study of using IIM images to inverse the lunar elements.

  4. Effect of Launaea procumbens extract on oxidative marker, p53, and CYP 2E1: a randomized control study

    Directory of Open Access Journals (Sweden)

    Rahmat Ali Khan

    2016-03-01

    Full Text Available Background: Ethyl acetate extracts of Launaea procumbens is used for the treatment of liver dysfunction as an herbal medicine in Pakistan. In this study, the protective effects of ethyl acetate extracts were evaluated against CCl4-induced liver injuries in rat. Methods: To examine the protective effects against oxidative stress of carbon tetrachloride in rats, 30 male rats were equally divided into 5 groups (6 rats. Among five groups, one was treated with CCl4 (3 ml/kg i.p. in olive oil b.w. twice a week for 4 weeks. Others were orally fed with extracts (100, 200 mg/kg b.w., with CCl4 twice a week for 4 weeks. Results: Administration of CCl4 altered the serum marker enzymes, lipid profile, CYP 2E1, p53 expression, antioxidant enzymes, nuclear organizer regions (AgNORs, and DNA. Supplement of L. procumbens ameliorated the effects of CCl4, improved CYP 2E1, p53, and increased the activities of antioxidant enzymes while activity of liver marker enzymes (ALP, ALT, AST, g-GT and contents of lipid per oxidation contents (TBARS, AgNORs, and DNA fragmentation were decreased. Similarly body weight was increased while liver and relative liver weight was decreased with co-administration of various extracts, suggesting that L. procumbens effectively protect liver against the CCl4-induced oxidative damage in rats. Conclusion: The hepatoprotective and free radical scavenging effects might be due to the presence of bioactive constituents in the extract.

  5. Purification and characterization of a potential antifungal protein from Bacillus subtilis E1R-J against Valsa mali.

    Science.gov (United States)

    Wang, N N; Yan, X; Gao, X N; Niu, H J; Kang, Z S; Huang, L L

    2016-04-01

    In order to identify the antagonistic substances produced by Bacillus subtilis E1R-J as candidate of biocontrol agents for controlling Apple Valsa Canker, hydrochloric acid precipitation, reverse phase chromatography, gel filtration, and ion exchange chromatography were used. The purified fraction EP-2 showed a single band in native-polyacrylamide gel electrophoresis (native-PAGE) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Fraction EP-2 was eluted from native-PAGE and showed a clear inhibition zone against V. mali 03-8. These results prove that EP-2 is one of the most important antifungal substances produced by B. subtilis E1R-J in fermentation broth. SDS-PAGE and Nano-LC-ESI-MS/MS analysis results demonstrated that EP-2 was likely an antifungal peptide (trA0A086WXP9), with a relative molecular mass of 12.44 kDa and isoelectric point of 9.94. The examination of antagonistic mechanism under SEM and TEM showed that EP-2 appeared to inhibit Valsa mali 03-8 by causing hyphal swelling, distortion, abnormality and protoplasts extravasation. Inhibition spectrum results showed that antifungal protein EP-2 had significantly inhibition on sixteen kinds of plant pathogenic fungi. The stability test results showed that protein EP-2 was stable with antifungal activity at temperatures as high as 100 °C for 30 min and in pH values ranging from 1.0 to 8.0, or incubated with each 5 mM Cu(2+), Zn(2+), Mg(2+), or K(+). However, the antifungal activity was negatively affected by Proteinase K treatment.

  6. Lunar iron and optical maturity mapping: Results from partial least squares modeling of Chang'E-1 IIM data

    Science.gov (United States)

    Sun, Lingzhi; Ling, Zongcheng; Zhang, Jiang; Li, Bo; Chen, Jian; Wu, Zhongchen; Liu, Jianzhong

    2016-12-01

    Iron and optical maturity (OMAT) are two key geological marks of the Moon that closely related to its geochemical evolution and interactions between surface and space environment. We apply Partial Least Squares (PLS) regression to Chang'E-1 Imaging Interferometer (IIM) (32 bands between 480 and 960 nm) in mapping lunar global FeO and OMAT, and the FeO and OMAT values are derived based on reasonable spectral parameters (absorbance, band ratios, TiO2 and maturity sensitive parameters, etc.). After been calibrated by the FeO map from Lunar Prospector Gamma-Ray Spectrometer (LP-GRS), the global FeO map derived from PLS modeling shows a quantitatively more reasonable result consistent with previous remote sensing results (LP) as well as lunar feldspathic meteorite studies and Chang'E-3 landing site. Based on the new FeO map by Chang'E-1, we discover a compositional inhomogeneity across lunar highland regions, which has not been suggested by previous datasets (e.g., Clementine UVVIS). Furthermore, we suggest that at least part of the FeO enrichments in highlands would be caused by mixing of highland and mare materials. The IIM derived OMAT map does not suggest a dichotomy of the lunar highlands and mare regions, implying the compositional differences between those two terrains have been suppressed. We further check the maturity effect for the young mare basalts (20 wt%) and ultrahigh-TiO2 (>10 wt%) tend to have greater OMAT values and vary little with ages; (3) this may be due to the distinct optical maturity effects of ultramafic minerals (i.e., ultrahigh Fe and Ti) and/or the spectral blue shifts of abundant ilmenite.

  7. Assessment of the Olea pollen and its major allergen Ole e 1 concentrations in the bioearosol of two biogeographical areas

    Science.gov (United States)

    Moreno-Grau, S.; Aira, M. J.; Elvira-Rendueles, B.; Fernández-González, M.; Fernández-González, D.; García-Sánchez, A.; Martínez-García, M. J.; Moreno, J. M.; Negral, L.; Vara, A.; Rodríguez-Rajo, F. J.

    2016-11-01

    The Olea pollen is currently an important allergy source. In some regions of Southern Spain, olive pollen is the main cause of allergic sensitization exceeding 40% of the sensitized individuals. Due to the scarce presence of olive trees in Northern Spain, limited to some cultivated fields in the South of the Galicia region where they also grow wild, only 8% of the sensitized individuals showed positive results for Olea pollen. The aim of the paper was to assess the behaviour pattern of the Olea pollen and its aeroallergens in the atmosphere, as this information could help us to improve the understanding and prevention of clinical symptoms. Airborne Olea pollen and Ole e 1 allergens were quantified in Cartagena (South-eastern Spain) and Ourense (North-western Spain). A volumetric pollen trap and a Burkard Cyclone sampler were used for pollen and allergen quantification. The Olea flowering took place in April or May in both biometeorological sampling areas. The higher concentrations were registered in the Southern area of Spain, for both pollen and Ole e 1, with values 8 times higher for pollen concentrations and 40 times higher for allergens. An alternate bearing pattern could be observed, characterized by years with high pollen values and low allergen concentrations and vice versa. Moreover, during some flowering seasons the allergen concentrations did not correspond to the atmospheric pollen values. Variations in weather conditions or Long Distance Transport (LDT) processes could explain the discordance. The back trajectory analysis shows that the most important contributions of pollen and allergens in the atmosphere are coincident with air masses passing through potential source areas. The exposure to olive pollen may not be synonym of antigen exposure.

  8. Native Folding of a Recombinant gpE1/gpE2 Heterodimer Vaccine Antigen from a Precursor Protein Fused with Fc IgG.

    Science.gov (United States)

    Logan, Michael; Law, John; Wong, Jason Alexander Ji-Xhin; Hockman, Darren; Landi, Amir; Chen, Chao; Crawford, Kevin; Kundu, Juthika; Baldwin, Lesley; Johnson, Janelle; Dahiya, Anita; LaChance, Gerald; Marcotrigiano, Joseph; Law, Mansun; Foung, Steven; Tyrrell, Lorne; Houghton, Michael

    2017-01-01

    A recombinant strain HCV1 (hepatitis C virus [HCV] genotype 1a) gpE1/gpE2 (E1E2) vaccine candidate was previously shown by our group to protect chimpanzees and generate broad cross-neutralizing antibodies in animals and humans. In addition, recent independent studies have highlighted the importance of conserved neutralizing epitopes in HCV vaccine development that map to antigenic clusters in E2 or the E1E2 heterodimer. E1E2 can be purified using Galanthis nivalis lectin agarose (GNA), but this technique is suboptimal for global production. Our goal was to investigate a high-affinity and scalable method for isolating E1E2. We generated an Fc tag-derived (Fc-d) E1E2 that was selectively captured by protein G Sepharose, with the tag being removed subsequently using PreScission protease. Surprisingly, despite the presence of the large Fc tag, Fc-d E1E2 formed heterodimers similar to those formed by GNA-purified wild-type (WT) E1E2 and exhibited nearly identical binding profiles to HCV monoclonal antibodies that target conserved neutralizing epitopes in E2 (HC33.4, HC84.26, and AR3B) and the E1E2 heterodimer (AR4A and AR5A). Antisera from immunized mice showed that Fc-d E1E2 elicited anti-E2 antibody titers and neutralization of HCV pseudotype viruses similar to those with WT E1E2. Competition enzyme-linked immunosorbent assays (ELISAs) showed that antisera from immunized mice inhibited monoclonal antibody binding to neutralizing epitopes. Antisera from Fc-d E1E2-immunized mice exhibited stronger competition for AR3B and AR5A than the WT, whereas the levels of competition for HC84.26 and AR4A were similar. We anticipate that Fc-d E1E2 will provide a scalable purification and manufacturing process using protein A/G-based chromatography.

  9. State-dependent electrostatic interactions of S4 arginines with E1 in S2 during Kv7.1 activation.

    Science.gov (United States)

    Wu, Dick; Delaloye, Kelli; Zaydman, Mark A; Nekouzadeh, Ali; Rudy, Yoram; Cui, Jianmin

    2010-06-01

    The voltage-sensing domain of voltage-gated channels is comprised of four transmembrane helices (S1-S4), with conserved positively charged residues in S4 moving across the membrane in response to changes in transmembrane voltage. Although it has been shown that positive charges in S4 interact with negative countercharges in S2 and S3 to facilitate protein maturation, how these electrostatic interactions participate in channel gating remains unclear. We studied a mutation in Kv7.1 (also known as KCNQ1 or KvLQT1) channels associated with long QT syndrome (E1K in S2) and found that reversal of the charge at E1 eliminates macroscopic current without inhibiting protein trafficking to the membrane. Pairing E1R with individual charge reversal mutations of arginines in S4 (R1-R4) can restore current, demonstrating that R1-R4 interact with E1. After mutating E1 to cysteine, we probed E1C with charged methanethiosulfonate (MTS) reagents. MTS reagents could not modify E1C in the absence of KCNE1. With KCNE1, (2-sulfonatoethyl) MTS (MTSES)(-) could modify E1C, but [2-(trimethylammonium)ethyl] MTS (MTSET)(+) could not, confirming the presence of a positively charged environment around E1C that allows approach by MTSES(-) but repels MTSET(+). We could change the local electrostatic environment of E1C by making charge reversal and/or neutralization mutations of R1 and R4, such that MTSET(+) modified these constructs depending on activation states of the voltage sensor. Our results confirm the interaction between E1 and the fourth arginine in S4 (R4) predicted from open-state crystal structures of Kv channels and reveal an E1-R1 interaction in the resting state. Thus, E1 engages in electrostatic interactions with arginines in S4 sequentially during the gating movement of S4. These electrostatic interactions contribute energetically to voltage-dependent gating and are important in setting the limits for S4 movement.

  10. Antibody to E1 peptide of hepatitis C virus genotype 4 inhibits virus binding and entry to HepG2 cells in vitro

    Science.gov (United States)

    EL-Awady, Mostafa K; Tabll, Ashraf A; Atef, Khaled; Yousef, Samar S; Omran, Moataza H; El-Abd, Yasmin; Bader-Eldin, Noha G; Salem, Ahmad M; Zohny, Samir F; El-Garf, Wael T

    2006-01-01

    AIM: To analyze the neutralizing activity of antibodies against E1 region of hepatitis C virus (HCV). Specific polyclonal antibody was raised via immunization of New Zealand rabbits with a synthetic peptide that had been derived from the E1 region of HCV and was shown to be highly conserved among HCV published genotypes. METHODS: Hyper-immune HCV E1 antibodies were incubated over night at 4 °C with serum samples positive for HCV RNA, with viral loads ranging from 615 to 3.2 million IU/ mL. Treated sera were incubated with HepG2 cells for 90 min. Blocking of viral binding and entry into cells by anti E1 antibody were tested by means of RT-PCR and flow cytometry. RESULTS: Direct immunostaining using FITC conjugated E1 antibody followed by Flow cytometric analysis showed reduced mean fluorescence intensity in samples pre-incubated with E1 antibody compared with untreated samples. Furthermore, 13 out of 18 positive sera (72%) showed complete inhibition of infectivity as detected by RT-PCR. CONCLUSION: In house produced E1 antibody, blocks binding and entry of HCV virion infection to target cells suggesting the involvement of this epitope in virus binding and entry. Isolation of these antibodies that block virus attachment to human cells are useful as therapeutic reagents. PMID:16688798

  11. Antibody to E1 peptide of hepatitis C virus genotype 4 inhibits virus binding and entry to HepG2 cells in vitro

    Institute of Scientific and Technical Information of China (English)

    Mostafa K EL-Awady; Wael T El-Garf; Ashraf A Tabll; Khaled Atef; Samar S Yousef; Moataza H Omran; Yasmin El-Abd; Noha G Bader-Eldin; Ahmad M Salem; Samir F Zohny

    2006-01-01

    AIM: To analyze the neutralizing activity of antibodies against E1 region of hepatitis C virus (HCV). Specific polyclonal antibody was raised via immunization of New Zealand rabbits with a synthetic peptide that had been derived from the E1 region of HCV and was shown to be highly conserved among HCV published genotypes.METHODS: Hyper-immune HCV E1 antibodies were incubatecd over night at 4 ℃ with serum samples positive for HCV RNA, with viral loads ranging from 615 to 3.2 million IU/mL. Treated sera were incubated with HepG2 cells for 90 min. Blocking of viral binding and entry into cells by anti E1 antibody were tested by means of RTPCR and flow cytometry.RESULTS: Direct immunostaining using FTTC conjugated E1 antibody followed by Flow cytometric analysis showed reduced mean fluorescence intensity in samples pre-incubated with E1 antibody compared with untreated samples. Furthermore, 13 out of 18 positive sera (72%)showed complete inhibition of infectivity as detected by RT-PCR.CONCLUSION: In house produced E1 antibody, blocks binding and entry of HCV virion infection to target cells suggesting the involvement of this epitope in virus binding and entry. Isolation of these antibodies that block virus attachment to human cells are useful as therapeutic reagents.

  12. [Screen potential CYP450 2E1 inhibitors from Chinese herbal medicine based on support vector regression and molecular docking method].

    Science.gov (United States)

    Chen, Xi; Lu, Fang; Jiang, Lu-di; Cai, Yi-Lian; Li, Gong-Yu; Zhang, Yan-Ling

    2016-07-01

    Inhibition of cytochrome P450 (CYP450) enzymes is the most common reasons for drug interactions, so the study on early prediction of CYPs inhibitors can help to decrease the incidence of adverse reactions caused by drug interactions.CYP450 2E1(CYP2E1), as a key role in drug metabolism process, has broad spectrum of drug metabolism substrate. In this study, 32 CYP2E1 inhibitors were collected for the construction of support vector regression (SVR) model. The test set data were used to verify CYP2E1 quantitative models and obtain the optimal prediction model of CYP2E1 inhibitor. Meanwhile, one molecular docking program, CDOCKER, was utilized to analyze the interaction pattern between positive compounds and active pocket to establish the optimal screening model of CYP2E1 inhibitors.SVR model and molecular docking prediction model were combined to screen traditional Chinese medicine database (TCMD), which could improve the calculation efficiency and prediction accuracy. 6 376 traditional Chinese medicine (TCM) compounds predicted by SVR model were obtained, and in further verification by using molecular docking model, 247 TCM compounds with potential inhibitory activities against CYP2E1 were finally retained. Some of them have been verified by experiments. The results demonstrated that this study could provide guidance for the virtual screening of CYP450 inhibitors and the prediction of CYPs-mediated DDIs, and also provide references for clinical rational drug use. Copyright© by the Chinese Pharmaceutical Association.

  13. Possible Mechanisms of Ethanol-Mediated Colorectal Carcinogenesis: The Role of Cytochrome P4502E1, Etheno-DNA Adducts, and the Anti-Apoptotic Protein Mcl-1.

    Science.gov (United States)

    Koehler, Bruno Christian; Arslic-Schmitt, Tatjana; Peccerella, Theresa; Scherr, Anna-Lena; Schulze-Bergkamen, Henning; Bruckner, Thomas; Gdynia, Georg; Jäger, Dirk; Mueller, Sebastian; Bartsch, Helmut; Seitz, Helmut K

    2016-10-01

    Chronic alcohol consumption is a risk factor for colorectal cancer. The mechanisms by which ethanol (EtOH) exerts its carcinogenic effect on the colorectal mucosa are not clear and may include oxidative stress with the action of reactive oxygen species (ROS) generated through EtOH metabolism via cytochrome P4502E1 (CYP2E1) leading to carcinogenic etheno-DNA adducts. ROS may also induce apoptosis. However, the effect of chronic EtOH consumption on CYP2E1, etheno-DNA adducts as well as anti-apoptotic proteins in the colorectal mucosa of heavy drinkers without colorectal inflammation is still not known. Rectal biopsies from 32 alcoholics (>60 g EtOH/d) and from 12 controls (alcoholics and control patients. However, CYP2E1 and etheno-DNA adducts correlated significantly when both groups were combined (p colorectal CYP2E1 was not found to be significantly increased in alcoholics, CYP2E1 correlated overall with the level of etheno-DNA adducts in the colorectal mucosa, which identifies CYP2E1 as an important factor in colorectal carcinogenesis. Most importantly, however, is the up-regulation of the anti-apoptotic protein Mcl-1 in heavy drinkers counteracting apoptosis and possibly stimulating cancer development. Copyright © 2016 by the Research Society on Alcoholism.

  14. Heterologous Acidothermus cellulolyticus 1,4-β-Endoglucanase E1 Produced Within the Corn Biomass Converts Corn Stover Into Glucose

    Science.gov (United States)

    Ransom, Callista; Balan, Venkatesh; Biswas, Gadab; Dale, Bruce; Crockett, Elaine; Sticklen, Mariam

    Commercial conversion of lignocellulosic biomass to fermentable sugars requires inexpensive bulk production of biologically active cellulase enzymes, which might be achieved through direct production of these enzymes within the biomass crops. Transgenic corn plants containing the catalytic domain of Acidothermus cellulolyticus E1 endo-1,4-β glucanase and the bar bialaphos resistance coding sequences were generated after Biolistic® (BioRad Hercules, CA) bombardment of immature embryo-derived cells. E1 sequences were regulated under the control of the cauliflower mosaic virus 35S promoter and tobacco mosaic virus translational enhancer, and E1 protein was targeted to the apoplast using the signal peptide of tobacco pathogenesis-related protein to achieve accumulation of this enzyme. The integration, expression, and segregation of E1 and bar transgenes were demonstrated, respectively, through Southern and Western blotting, and progeny analyses. Accumulation of up to 1.13% of transgenic plant total soluble proteins was detected as biologically active E1 by enzymatic activity assay. The corn-produced, heterologous E1 could successfully convert ammonia fiber explosion-pretreated corn stover polysaccharides into glucose as a fermentable sugar for ethanol production, confirming that the E1 enzyme is produced in its active from.

  15. E4orf6 variants with separate abilities to augment adenovirus replication and direct nuclear localization of the E1B 55-kilodalton protein.

    Science.gov (United States)

    Orlando, Joseph S; Ornelles, David A

    2002-02-01

    The E4orf6 protein of group C adenovirus is an oncoprotein that, in association with the E1B 55-kDa protein and by E1B-independent means, promotes virus replication. An arginine-faced amphipathic alpha-helix in the E4orf6 protein is required for the E4orf6 protein to direct nuclear localization of the E1B 55-kDa protein and to enhance replication of an E4 deletion virus. In this study, E4orf6 protein variants containing arginine substitutions in the amphipathic alpha-helix were analyzed. Two of the six arginine residues within the alpha-helix, arginine-241 and arginine-243, were critical for directing nuclear localization of the E1B 55-kDa protein. The four remaining arginine residues appear to provide a net positive charge for the E4orf6 protein to direct nuclear localization of the E1B 55-kDa protein. The molecular determinants of the arginine-faced amphipathic alpha-helix that were required for the functional interaction between the E4orf6 and E1B 55-kDa proteins seen in the transfected cell differed from those required to support a productive infection. Several E4orf6 protein variants with arginine-to-glutamic acid substitutions that failed to direct nuclear localization of the E1B 55-kDa protein restored replication of an E4 deletion virus. Additionally, a variant containing an arginine-to-alanine substitution at position 243 that directed nuclear localization of the E1B 55-kDa protein failed to enhance virus replication. These results indicate that the ability of the E4orf6 protein to relocalize the E1B 55-kDa protein to the nucleus can be separated from the ability of the E4orf6 protein to support a productive infection.

  16. Recombinant protein of tissue inhibitor of metaIloproteinase-3 induces apoptosis of mouse MC3T3-E1 osteoblasts

    Institute of Scientific and Technical Information of China (English)

    袁凌青

    2006-01-01

    Objective To investigate the action of recombinantprotein of tissue inhibitor of metalloproteinase-3 (TIMP-3) on apoptosis of MC3T3-E1 osteoblasts. Methods Cell survival rate and apoptosis were measured by MTT and ELISA respectively. The expressions of Fas, FasL, Bel-2, Bax, caspase-3 , caspase-8, cytochrome c and phosphorylations of JNK, p38 and extracellular signalregulated kinase (ERK) 1/2 were analysed by Western blotting. Results TIMP-3 reduced survival rate of MC3T3-E1 cells and promoted apoptosis of MC3T3-E1

  17. E1 Nino发生相联系的热带大气系统动能的变化

    Institute of Scientific and Technical Information of China (English)

    李崇银; 李桂龙

    1995-01-01

    @@ 作者之一首先在1988年指出E1 Nino事件的前期征兆在赤道西太平洋,其后的进一步研究更明确指出,东亚冬季风异常与E1 Nino有相互作用[1,2].通过资料分析和GCM的数值模拟,我们最近又指出热带大气季节内振荡与E1 Nino也存在明显的相互作用[3].

  18. Effect of pollen from Typha angustata on hydrogen peroxide induced toxicity in osteoblastic MC3T3-E1 cells.

    Science.gov (United States)

    Lee, Young Soon; Choi, Eun Mi

    2012-02-01

    Typha angustata is a traditional Chinese medicine, commonly used for a variety of clinical disorders, including atherosclerosis and wound healing. In the present study, the protective effects of T. angustata pollen extract (TE) on the response of osteoblast to oxidative stress were evaluated. Osteoblastic MC3T3-E1 cells were incubated with H(2)O(2) and/or TE, and markers of osteoblast function and oxidative damage were examined. TE treatment significantly (P < 0.05) reversed the cytotoxic effect of H(2)O(2) and this effect was blocked by ICI182780, suggesting that TE's effect might be partly involved in estrogen action. TE significantly (P < 0.05) increased collagen content, alkaline phosphatase activity, calcium deposition of osteoblasts in the presence of H(2)O(2) and these effects were blocked by rottlerin and PD98059, suggesting that the induction of differentiation by TE is associated with increased activation of protein kinase C and ERK. Moreover, H(2)O(2)-induced reduction of osteocalcin was significantly recovered in the presence of TE. Pretreatment with TE also decreased the increase in receptor activator of nuclear factor-kB ligand, malondialdehyde, and protein carbonyl induced by H(2)O(2). These results suggest that the pollen of T. angustata may be useful for the protection of H(2)O(2)-induced oxidative damage and dysfunction in osteoblasts. © 2011 John Wiley & Sons A/S.

  19. The effects of a special Agnus castus extract (BP1095E1) on prolactin secretion in healthy male subjects.

    Science.gov (United States)

    Merz, P G; Gorkow, C; Schrödter, A; Rietbrock, S; Sieder, C; Loew, D; Dericks-Tan, J S; Taubert, H D

    1996-01-01

    The effects of three doses of a special Agnus castus extract (BP1095E1)--extracts from 120 mg, 240 mg and 480 mg of drug per day--were examined within the framework of a placebo-controlled clinical study of tolerance and prolactin secretion in 20 healthy male subjects during a period of 14 days. There was good tolerance during the study as regards the following: adverse effects, the effects on blood pressure and heart rate, blood count, Quick's test, clinical chemistry as well as testosterone, FSH and LH values. During each study phase the 24-hour prolactin secretion profile was measured from the penultimate to the final day, and the amount of prolactin release was monitored an hour after TRH stimulation on the last day. A significant increase in the 24-hour profile was registered with the lowest dose in comparison to placebo, the opposite being the case with the higher doses, i.e. a slight reduction. In contrast to the administration of placebo, the 1-hour AUC after TRH stimulation resulted in a significant increase with the lowest dose and a significant reduction with the highest dose. The results suggest effects of the special Agnus castus extract which are dependent on the dose administered and the initial level of prolactin concentration.

  20. Epicteto: Diatribes 1.29 e 1.30 - uma síntese da filosofia epictetiana das Diatribes

    Directory of Open Access Journals (Sweden)

    Aldo Dinucci

    2015-04-01

    Full Text Available As diatribes 1.29 e 1.30 encerram o livro primeiro das Diatribes de Epicteto e nos oferecem uma síntese da filosofia de Epicteto. Em 1.29.1, Epicteto nos diz que a essência (ousia do bem do ser humano “é uma capacidade de escolha de certa qualidade”, isto é, a escolha disposta de certa maneira. Por essa definição, todas as demais coisas que se encontram fora do âmbito da escolha e, consequentemente, do bem humano são somente materiais para a escolha (1.29.2, a partir dos quais ela construirá seu próprio bem ou seu próprio mal. Temos aí posta de outra forma a distinção entre coisas sob nosso encargo e coisas que não estão sob nosso encargo (eph’hemin kai ouk eph’hemin, sobre qual Epicteto trata detidamente em diatribe 1.1 e no primeiro capítulo do Encheirídion

  1. Quantitative Crystal Structure Analysis of (E-1-[(2-Chloro-1,3-thiazol-5-ylmethyl]-3-methyl-2-nitroguanidine

    Directory of Open Access Journals (Sweden)

    Dhananjay Dey

    2014-01-01

    Full Text Available The crystal structure of a biologically active (E-1-[(2-chloro-1,3-thiazol-5-ylmethyl]-3-methyl-2-nitroguanidine with molecular formula C6H8N5O2ClS has been investigated based on the molecular conformation and the supramolecular packing in terms of intermolecular interactions involving N–H⋯O, N–H⋯N, and C–H⋯O–N (nitro group, C–H⋯N (thiazol hydrogen bonds, offset π–π stacking, C–H⋯π and N(–NO2⋯C=N intermolecular interactions. Furthermore, a short C–Cl⋯O–N contact is also present which contributes towards the crystal packing. The lattice energy of the title compound has been calculated using the PIXEL approach (the Coulomb-London-Pauli (CLP model and compared with periodic calculations performed using CRYSTAL09. In addition, Hirshfeld surface analysis and fingerprint plots provide a platform for the evaluation of the contribution of different intermolecular interactions towards the packing behaviour.

  2. Traf2 interacts with Smad4 and regulates BMP signaling pathway in MC3T3-E1 osteoblasts

    Energy Technology Data Exchange (ETDEWEB)

    Shimada, Koichi, E-mail: shimada-ki@dent.nihon-u.ac.jp [Department of Periodontology, Nihon University School of Dentistry, Tokyo (Japan); Division of Advanced Dental Treatment, Dental Research Center, Nihon University School of Dentistry, Tokyo (Japan); Ikeda, Kyoko [Department of Periodontology, Nihon University School of Dentistry, Tokyo (Japan); Ito, Koichi [Department of Periodontology, Nihon University School of Dentistry, Tokyo (Japan); Division of Advanced Dental Treatment, Dental Research Center, Nihon University School of Dentistry, Tokyo (Japan)

    2009-12-18

    Bone morphogenetic proteins (BMPs) play important roles in osteoblast differentiation and maturation. In mammals, the BMP-induced receptor-regulated Smads form complexes with Smad4. These complexes translocate and accumulate in the nucleus, where they regulate the transcription of various target genes. However, the function of Smad4 remains unclear. We performed a yeast two-hybrid screen using Smad4 as bait and a cDNA library derived from bone marrow, to indentify the proteins interacting with Smad4. cDNA clones for Tumor necrosis factor (TNF) receptor-associated factor 2 (Traf2) were identified, and the interaction between the endogenous proteins was confirmed in the mouse osteoblast cell line MC3T3-E1. To investigate the function of Traf2, we silenced it with siRNA. The level of BMP-2 protein in the medium, the expression levels of the Bmp2 gene and BMP-induced transcription factor genes, including Runx2, Dlx5, Msx2, and Sp7, and the phosphorylated-Smad1 protein level were increased in cells transfected with Traf2 siRNA. The nuclear accumulation of Smad1 increased with TNF-{alpha} stimulation for 30 min at Traf2 silencing. These results suggest that the TNF-{alpha}-stimulated nuclear accumulation of Smad1 may be dependent on Traf2. Thus, the interaction between Traf2 and Smad4 may play a role in the cross-talk between TNF-{alpha} and BMP signaling pathways.

  3. MC3T3-E1 cell response to stainless steel 316L with different surface treatments.

    Science.gov (United States)

    Zhang, Hongyu; Han, Jianmin; Sun, Yulong; Huang, Yongling; Zhou, Ming

    2015-11-01

    In the present study, stainless steel 316L samples with polishing, aluminum oxide blasting, and hydroxyapatite (HA) coating were prepared and characterized through a scanning electron microscope (SEM), optical interferometer (surface roughness, Sq), contact angle, surface composition and phase composition analyses. Osteoblast-like MC3T3-E1 cell adhesion on the samples was investigated by cell morphology using a SEM (4h, 1d, 3d, 7d), and cell proliferation was assessed by MTT method at 1d, 3d, and 7d. In addition, adsorption of bovine serum albumin on the samples was evaluated at 1h. The polished sample was smooth (Sq: 1.8nm), and the blasted and HA coated samples were much rougher (Sq: 3.2μm and 7.8μm). Within 1d of incubation, the HA coated samples showed the best cell morphology (e.g., flattened shape and complete spread), but there was no significant difference after 3d and 7d of incubation for all the samples. The absorbance value for the HA coated samples was the highest after 1d and 3d of incubation, indicating better cell viability. However, it reduced to the lowest value at 7d. Protein adsorption on the HA coated samples was the highest at 1h. The results indicate that rough stainless steel surface improves cell adhesion and morphology, and HA coating contributes to superior cell adhesion, but inhibits cell proliferation.

  4. Abundance and distribution of radioelements in lunar terranes: Results of Chang'E-1 gamma ray spectrometer data

    Science.gov (United States)

    Chen, Jian; Ling, Zongcheng; Li, Bo; Zhang, Jiang; Sun, Lingzhi; Liu, Jianzhong

    2016-02-01

    The gamma ray spectrometer (GRS) onboard Chang'E-1 has acquired valuable datasets recording the gamma ray intensities from radioelements (Potassium (K), Thorium (Th) and Uranium (U), etc.) on lunar surface. We extracted the elemental concentrations from the GRS data with spectral fitting techniques and mapped the global absolute abundance of radioelements in terms of the ground truths from lunar samples and meteorites. The obtained global concentration maps of these radioelements indicate heterogeneous distribution among three major lunar crustal terranes (i.e., Procellarum KREEP Terrane (PKT), Feldspathic Highlands Terrane (FHT), and South Pole Aitken Terrane (SPAT)) in relation with their origin and distinct geologic history. The majority of radioelements are restricted in PKT, approving the scenario of KREEP (Potassium (K), rare earth elements (REE), Phosphorus (P)) residua concentrating under the Procellarum region. Moreover, we found the consistency of distribution for radioelements and basalts, concluding that the subsequent volcanism might be associated with local concentrations of radioelements in western Oceanus Procellarum and northwestern South Pole Aitken Basin. The prominent and asymmetric radioactive signatures were confirmed in SPAT comparing to FHT dominated by low level radioactivity, while the magnitudes are much lower than that of PKT, indicating a primary geochemical heterogeneity for the Moon.

  5. Aurantio-obtusin stimulates chemotactic migration and differentiation of MC3T3-E1 osteoblast cells.

    Science.gov (United States)

    Vishnuprasad, Chethala N; Tsuchiya, Tomoko; Kanegasaki, Shiro; Kim, Joon Ho; Han, Sung Soo

    2014-05-01

    Osteoporosis is one of the major metabolic bone diseases and is among the most challenging noncommunicable diseases to treat. Although there is an increasing interest in identifying bioactive molecules for the prevention and management of osteoporosis, such studies principally focus only on differentiation and mineralization of osteoblasts or inhibition of osteoclast activity. Stimulation of osteoblast migration must be a promising osteoanabolic strategy for improved metabolic bone disease therapy. In this study, we show that an anthraquinone derivative, aurantio-obtusin, stimulated chemotactic migration of MC3T3-E1 osteoblast cells in a concentration-dependent manner. The use of a real-time chemotaxis analyzing system, TAXIScan, facilitated the evaluation of both velocity and directionality of osteoblast migration in response to the compound. Besides migration, the compound stimulated osteoblast differentiation and mineralization. Taken together, the data presented in this paper demonstrate that aurantio-obtusin is a promising osteoanabolic compound of natural origin with potential therapeutic applications in the prevention of osteoporosis and other metabolic bone diseases.

  6. Significance of phentolamine redosing during prostaglandin E1 penile color Doppler ultrasonography in diagnosis of vascular erectile dysfunction.

    Science.gov (United States)

    Arafa, Mohamed; Eid, Hazem; Shamloul, Rany

    2007-05-01

    Recently, it was reported that phentolamine redosing during penile duplex can abolish a false diagnosis of venous leakage in patients with impotence. The aim of this study is to identify any useful role of phentolamine redosing in diagnosis of venogenic impotence. Sixty-seven consecutive patients complaining of weak erection for at least 6 months were included in this study. Penile color Doppler ultrasound (CDU) was performed using a 7.5 MHz linear array transducer with a color flow mapping capability. Following intracavernous injection of 20 microg prostaglandin E1 (PGE1), all patients with persistent end diastolic velocity (EDV) >5 cm/sec with an erectile response of E3 or lower, 20 min after intracavernosal injection of PGE1, were asked to revisit our clinic for a second CDU, 2 weeks later. During initial CDU examination, all 67 patients experienced poor response to 20 microg PGE1 with their average peak systolic velocity (PSV) and EDV being 42.8 and 6.6 cm/sec, respectively. The second CDU examination had similar results to the first one. Addition of 2 mg phentolamine did not significantly change the PSV and EDV of cavernosal arteries in any of the 67 patients. In conclusion, addition of intracavernous phentolamine during PGE1 CDU examination carries no advantage over the use of PGE1 alone regarding cavernosal artery response in patients with suspected venogenic EDV.

  7. Crystal structure, spectroscopic characterization and density functional studies of (E)-1-((3-methoxyphenylimino)methyl)naphthalen-2-ol

    Science.gov (United States)

    Alpaslan, Gökhan; Macit, Mustafa

    2014-03-01

    The Schiff base compound (E)-1-((3-methoxyphenylimino)methyl)naphthalen-2-ol was synthesized from the reaction of 2-hydroxy-1-naphthaldehyde with 3-methoxyaniline. The structural properties of the compound has been characterized by using FT-IR, UV-vis and X-ray single-crystal methods. According to X-ray diffraction result, the title compound exists in the phenol-imine tautomeric form. The molecular geometry, vibrational frequencies of the compound in the ground state have been calculated using the density functional theory (DFT/B3LYP) method with the 6-311++G(d,p) basis set, and compared with the experimental data. The obtained results show that the optimized molecular geometry is well reproduce the crystal structure. The theoretical vibrational frequencies are in good agreement with the experimental values. The calculations of electronic absorption spectra of tautomeric forms of the compound were performed by using TD-DFT calculations both in the gas phase and ethanol solvent. To investigate the tautomeric stability, optimization calculations at the B3LYP/6-311++G(d,p) level were performed for the phenol-imine and keto-amine forms of the compound. According to calculated results, the OH form is more stable than NH form. In addition, molecular electrostatic potential (MEP), frontier molecular orbital analysis (HOMO-LUMO), thermodynamic and, non-linear optical (NLO) properties of the compound were investigated using same theoretical calculations.

  8. Example-based super-resolution for single-image analysis from the Chang'e-1 Mission

    Science.gov (United States)

    Wu, Fan-Lu; Wang, Xiang-Jun

    2016-11-01

    Due to the low spatial resolution of images taken from the Chang'e-1 (CE-1) orbiter, the details of the lunar surface are blurred and lost. Considering the limited spatial resolution of image data obtained by a CCD camera on CE-1, an example-based super-resolution (SR) algorithm is employed to obtain high-resolution (HR) images. SR reconstruction is important for the application of image data to increase the resolution of images. In this article, a novel example-based algorithm is proposed to implement SR reconstruction by single-image analysis, and the computational cost is reduced compared to other example-based SR methods. The results show that this method can enhance the resolution of images using SR and recover detailed information about the lunar surface. Thus it can be used for surveying HR terrain and geological features. Moreover, the algorithm is significant for the HR processing of remotely sensed images obtained by other imaging systems.

  9. Origin of the large positive magnetoresistance of G e1 -xM nx granular thin films

    Science.gov (United States)

    Wakabayashi, Yuki K.; Akiyama, Ryota; Takeda, Yukiharu; Horio, Masafumi; Shibata, Goro; Sakamoto, Shoya; Ban, Yoshisuke; Saitoh, Yuji; Yamagami, Hiroshi; Fujimori, Atsushi; Tanaka, Masaaki; Ohya, Shinobu

    2017-01-01

    G e1 -xM nx (GeMn) granular thin films are a unique and promising material for spintronic applications owing to their large positive magnetoresistance (MR). Previous studies of GeMn have suggested that the large MR is related to the nanospinodal decomposition of GeMn into Mn-rich ferromagnetic nanoparticles and a Mn-poor paramagnetic matrix. However, the microscopic origin of the MR has not yet been clarified. Here, we develop a method to separately investigate the magnetic properties of the nanoparticles and the matrix, utilizing the extremely high sensitivity of x-ray magnetic circular dichroism (XMCD) to the local magnetic state of each atom. We find that the MR ratio is proportional to the product of the magnetizations originating from the nanoparticles and the matrix. This result indicates that the spin-polarized holes in the nanoparticles penetrate into the matrix and that these holes undergo first order magnetic scattering by the paramagnetic Mn atoms in the matrix, which induces the large MR.

  10. The E1 beta-subunit of pyruvate dehydrogenase is surface-expressed in Lactobacillus plantarum and binds fibronectin.

    Science.gov (United States)

    Vastano, Valeria; Salzillo, Marzia; Siciliano, Rosa A; Muscariello, Lidia; Sacco, Margherita; Marasco, Rosangela

    2014-01-01

    Lactobacillus plantarum is among the species with a probiotic activity. Adhesion of probiotic bacteria to host tissues is an important principle for strain selection, because it represents a crucial step in the colonization process of either pathogens or commensals. Most bacterial adhesins are proteins, and a major target for them is fibronectin, an extracellular matrix glycoprotein. In this study we demonstrate that PDHB, a component of the pyruvate dehydrogenase complex, is a factor contributing to fibronectin-binding in L. plantarum LM3. By means of fibronectin overlay immunoblotting assay, we identified a L. plantarum LM3 surface protein with apparent molecular mass of 35 kDa. Mass spectrometric analysis shows that this protein is the pyruvate dehydrogenase E1 beta-subunit (PDHB). The corresponding pdhB gene is located in a 4-gene cluster encoding pyruvate dehydrogenase. In LM3-B1, carrying a null mutation in pdhB, the 35 kDa adhesin was not anymore detectable by immunoblotting assay. Nevertheless, the pdhB null mutation did not abolish pdhA, pdhC, and pdhD transcription in LM3-B1. By adhesion assays, we show that LM3-B1 cells bind to immobilized fibronectin less efficiently than wild type cells. Moreover, we show that pdhB expression is negatively regulated by the CcpA protein and is induced by bile.

  11. Pharmacodynamic comparison of prostaglandin E1 administered by different routes to rats%前列腺素E1经不同途径给药后的大鼠体内药效学比较

    Institute of Scientific and Technical Information of China (English)

    谷福根; 崔福德; 高永良

    2007-01-01

    本文研究了前列腺素E1(PGE1)分别经不同途径给药后的大鼠体内药效学,旨在寻找目前PGE1注射给药的替代途径.以PGE1降压效应作为药效学指标,以静脉注射为对照,分别测定PGE1经鼻腔、舌下、肌肉(im)、腹腔(ip)给药后的药效学参数,包括峰效应时间(Tmax),血压下降最大百分数(Emax,%),效应持续时间(Td)以及血压下降百分数-时间曲线下面积(AUC,%·min).研究结果表明,PGE1经上述途径给药后,药效学参数Emax,Td,AUC等均随给药剂量的增加而增大,提示存在明显的剂量-效应关系.根据所测Tmax值,推断上述给药途径其吸收速率的大小顺序为:鼻腔≈im>ip>舌下;依据所测药理生物利用度(PF)值,预测药物绝对生物利用度的顺序为:鼻腔>im≈ip>舌下.上述研究结果提示,PGE1经鼻腔与舌下黏膜给药,有望替代目前的注射给药.%The pharmacodynamics of prostaglandin E1 (PGE1) administered by different routes to rats was investigated in this paper. The hypotensive effect of PGE1 was used as an index of drug efficacy, pharmacodynamic parameters such as time to reach peak effect (Tmax), maximal percentage of blood pressure decrease (Emax, %), duration of effect (Td), and the area determined after PGE1 given to rats intranasally, sublingually, intraperitoneally (ip),and intramuscularly (im), separately, and compared with those obtained from intravenous (iv) administration. Similar to iv route, the pharmacodynamic parameters of PGE1 from the other administration routes, Emax, Td and in particular AUC values were all increased with increasing doses, showing dose-efficacy relationship. Tmax was found to be approximately 3-4 min for nasal route, 3-8 min for im, 6-8 min for ip and 12-30 min for sublingual route, separately. Thus, the order of magnitude of absorption rate of the drug was as follows: nasal≈im>ip>sublingual. If the pharmacological bioavailability (PF) for each administration route was

  12. Highly stereoselective synthesis of (Z,E)-1-halo-1,3-dienol esters via rearrangement of Fischer chromium chloro-carbenes using microwave irradiation.

    Science.gov (United States)

    Kashinath, Dhurke; Mioskowski, Charles; Falck, J R; Goli, Mohan; Meunier, Stéphane; Baati, Rachid; Wagner, Alain

    2009-05-07

    Functionalized (Z,E)-1-halo-1,3-dienol esters are synthesized in a highly stereoselective manner via CrCl2-mediated rearrangement of allylic trihalomethylcarbinol esters induced by microwave irradiation.

  13. New Views of Lunar Compositions and Rock Types Revealed by Spectroscopic Data from Chang'e-1 and Chang'e-3 Missions

    Science.gov (United States)

    Ling, Z. C.; Jolliff, B. L.

    2016-05-01

    Global hyperspectral imaging datasets of Chang'e-1 IIM have brought new science returns of lunar surface compositions and rock types. The in-situ spectroscopic measurements by Yutu yielded compositional constraints of a new type of basaltic rock.

  14. Preclinical pharmacology and toxicology study of Ad-hTERT-E1a-Apoptin, a novel dual cancer-specific oncolytic adenovirus

    Energy Technology Data Exchange (ETDEWEB)

    Qi, Yanxin [State Key Laboratory of Polymer Physics and Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022 (China); Institute of Military Veterinary, Academy of Military Medical Sciences of PLA, Changchun 130122 (China); Guo, Huanhuan [Institute of Military Veterinary, Academy of Military Medical Sciences of PLA, Changchun 130122 (China); Changchun Brother Biotech Co., Ltd., Changchun, 130000 (China); Hu, Ningning; He, Dongyun [Institute of Military Veterinary, Academy of Military Medical Sciences of PLA, Changchun 130122 (China); The Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Changchun 130122 (China); Zhang, Shi [Institute of Military Veterinary, Academy of Military Medical Sciences of PLA, Changchun 130122 (China); School of Clinical Medicine, Jilin University, Changchun 130001 (China); Chu, Yunjie [Affiliated Hospital of Changchun University of Traditional Chinese Medicine, Changchun 130021 (China); Huang, Yubin [State Key Laboratory of Polymer Physics and Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022 (China); Li, Xiao, E-mail: lixiao06@mails.jlu.edu.cn [Institute of Military Veterinary, Academy of Military Medical Sciences of PLA, Changchun 130122 (China); The Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Changchun 130122 (China); Sun, LiLi, E-mail: linjiaxiaoya@163.com [Department of Head and Neck Surgery, Tumor Hospital of Jilin Province, Changchun 130012 (China); Jin, Ningyi, E-mail: ningyij@126.com [Institute of Military Veterinary, Academy of Military Medical Sciences of PLA, Changchun 130122 (China); The Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Changchun 130122 (China)

    2014-10-15

    Clinical studies have demonstrated that conditionally replicating adenovirus is safe. We constructed an oncolytic adenovirus, Ad-hTERT-E1a-Apoptin, using a cancer-specific promoter (human telomerase reverse transcriptase promoter, hTERTp) and a cancer cell-selective apoptosis-inducing gene (Apoptin). Ad-hTERT-E1a-Apoptin was proven effective both in vitro and in vivo in our previous study. In this study, the preclinical safety profiles of Ad-hTERT-E1a-Apoptin in animal models were investigated. At doses of 5.0 × 10{sup 8}, 2.5 × 10{sup 9}, and 1.25 × 10{sup 10} viral particles (VP)/kg, Ad-hTERT-E1a-Apoptin had no adverse effects on mouse behavior, muscle cooperation, sedative effect, digestive system, and nervous systems, or on beagle cardiovascular and respiratory systems at 5.0 × 10{sup 8}, 2.5 × 10{sup 9}, and 1.25 × 10{sup 10} VP/kg doses. In acute toxicity tests in mice, the maximum tolerated dose > 5 × 10{sup 10} VP/kg. There was no inflammation or ulceration at the injection sites within two weeks. In repeat-dose toxicological studies, the no observable adverse effect levels of Ad-hTERT-E1a-Apoptin in rats (1.25 × 10{sup 10} VP/kg) and beagles (2.5 × 10{sup 9} VP/kg) were 62.5- and 12.5-fold of the proposed clinical dose, respectively. The anti-virus antibody was produced in animal sera. Bone marrow examination revealed no histopathological changes. Guinea pigs sensitized by three repeated intraperitoneal injections of 1.35 × 10{sup 10} VP/mL Ad-hTERT-E1a-Apoptin each and challenged by one intravenous injection of 1.67 × 10{sup 8} VP/kg Ad-hTERT-E1a-Apoptin did not exhibit any sign of systemic anaphylaxis. Our data from different animal models suggest that Ad-hTERT-E1a-Apoptin is a safe anti-tumor therapeutic agent. - Highlights: • We use the rodents and non-rodents animal models to evaluation Ad-hTERT-E1a-Apoptin. • Ad-hTERT-E1a-Apoptin is a safe anti-tumor therapeutic agent. • Demonstrate the safety and feasibility dose of injected Ad-hTERT-E

  15. Ethanol metabolism by alcohol dehydrogenase or cytochrome P450 2E1 differentially impairs hepatic protein trafficking and growth hormone signaling.

    Science.gov (United States)

    Doody, Erin E; Groebner, Jennifer L; Walker, Jetta R; Frizol, Brittnee M; Tuma, Dean J; Fernandez, David J; Tuma, Pamela L

    2017-09-01

    The liver metabolizes alcohol using alcohol dehydrogenase (ADH) and cytochrome P450 2E1 (CYP2E1). Both enzymes metabolize ethanol into acetaldehyde, but CYP2E1 activity also results in the production of reactive oxygen species (ROS) that promote oxidative stress. We have previously shown that microtubules are hyperacetylated in ethanol-treated polarized, hepatic WIF-B cells and livers from ethanol fed rats. We have also shown that enhanced protein acetylation correlates with impaired clathrin-mediated endocytosis, constitutive secretion and nuclear translocation and that the defects are likely mediated by acetaldehyde. However, the roles of CYP2E1-generated metabolites and ROS in microtubule acetylation and these alcohol-induced impairments have not been examined. To determine if CYP2E1-mediated alcohol metabolism is required for enhanced acetylation and the trafficking defects, we co-incubated cells with ethanol and diallyl sulfide (a CYP2E1 inhibitor) or N-acetyl cysteine (an anti-oxidant). Both agents failed to prevent microtubule hyperacetylation in ethanol-treated cells and also failed to prevent impaired secretion or clathrin-mediated endocytosis. Somewhat surprisingly, both NAS and DAC prevented impaired STAT5B nuclear translocation. Further examination of microtubule-independent steps of the pathway revealed that Jak2/STAT5B activation by growth hormone (GH) was prevented by DAS and NAC. These results were confirmed in ethanol-exposed HepG2 cells expressing only ADH or CYP2E1. Using quantitative RT-PCR, we further determined that ethanol exposure led to blunted GH-mediated gene expression. In conclusion, we determined that alcohol-induced microtubule acetylation and associated defects in microtubule-dependent trafficking are mediated by ADH metabolism whereas impaired microtubule-independent Jak2/STAT5B activation is mediated by CYP2E1 activity. Copyright © 2017, American Journal of Physiology-Gastrointestinal and Liver Physiology.

  16. Design and optimization of N-acylhydrazone pyrimidine derivatives as E. coli PDHc E1 inhibitors: Structure-activity relationship analysis, biological evaluation and molecular docking study.

    Science.gov (United States)

    He, Haifeng; Xia, Hongying; Xia, Qin; Ren, Yanliang; He, Hongwu

    2017-08-23

    By targeting the thiamin diphosphate (ThDP) binding site of Escherichia coli (E. coli) pyruvate dehydrogenase multienzyme complex E1 (PDHc E1), a series of novel 'open-chain' classes of ThDP analogs A, B, and C with N-acylhydrazone moieties was designed and synthesized to explore their activities against E. coli PHDc E1 in vitro and their inhibitory activity against microbial diseases were further evaluated in vivo. As a result, A1-23 exhibited moderate to potent inhibitory activities against E. coli PDHc E1 (IC50=0.15-23.55μM). The potent inhibitors A13, A14, A15, C2, had strong inhibitory activities with IC50 values of 0.60, 0.15, 0.39 and 0.34μM against E. coli PDHc E1 and with good enzyme-selective inhibition between microorganisms and mammals. Especially, the most powerful inhibitor A14 could 99.37% control Xanthimonas oryzae pv. Oryzae. Furthermore, the binding features of compound A14 within E. coli PDHc E1 were investigated to provide useful insights for the further construction of new inhibitor by molecular docking, site-directed mutagenesis, and enzymatic assays. The results indicated that A14 had most powerful inhibition against E. coli PDHc E1 due to the establishment of stronger interaction with Glu571, Met194, Glu522, Leu264 and Phe602 at active site of E.coli PDHc E1. It could be used as a lead compound for further optimization, and may have potential as a new microbicide. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Differences in Butadiene Adduct Formation between Rats and Mice Not Due to Selective Inhibition of CYP2E1 by Butadiene Metabolites

    Science.gov (United States)

    Pianalto, Kaila M.; Hartman, Jessica H.; Boysen, Gunnar; Miller, Grover P.

    2013-01-01

    CYP2E1 metabolizes 1,3-butadiene (BD) into genotoxic and possibly carcinogenic 1,2-epoxy-3-butene (EB), 1,2:3,4-diepoxybutane (DEB), and 1,2-epoxy-3,4-butanediol (EB-diol). The dose response of DNA and protein adducts derived from BD metabolites increase linearly at low BD exposures and then saturate at higher exposures in rats, but not mice. It was hypothesized that differences in adduct formation between rodents reflect more efficient BD oxidation in mice than rats. Herein, we assessed whether BD-derived metabolites selectively inhibit rat but not mouse CYP2E1 activity using B6C3F1 mouse and Fisher 344 rat liver microsomes. Basal CYP2E1 activities toward 4-nitrophenol were similar between rodents. Through IC50 studies, EB was the strongest inhibitor (IC50 54 μM, mouse; 98 μM, rat), BD-diol considerably weaker (IC50 1200 μM, mouse; 1000 μM, rat), and DEB inhibition nonexistent (IC50 >25 mM). Kinetic studies showed that in both species EB and BD-diol inhibited 4-nitrophenol oxidation through two-site mechanisms in which inhibition constants reflected trends observed in IC50 studies. None of the reactive epoxide metabolites inactivated CYP2E1 irreversibly. Thus, there was no selective inhibition or inactivation of rat CYP2E1 by BD metabolites relative to mouse Cyp2e1, and it can be inferred that CYP2E1 activity toward BD between rodent species would similarly not be impacted by the presence of BD metabolites. Inhibition of CYP2E1 by BD metabolites is then not responsible for the reported species difference in BD metabolism, formation of BD-derived DNA and protein adducts, mutagenicity and tumorigenesis. PMID:24021170

  18. Human liver estrone (E1), Estradiol (E2) and dehydroepiandrosterone (DHEA) sulfotransferases (STs): Comparison with thermostable (TS) and thermolabile (TL) phenol sulfotransferase (PST) activities

    Energy Technology Data Exchange (ETDEWEB)

    Hernandez, J.S.; Watson, R.W.G.; Weinshilboum, R.M. (Mayo Clinic/Mayo Foundation, Rochester, MN (United States))

    1991-03-11

    Sulfation plays an important role in the metabolism of E1, E2 and DHEA in humans. The relationship between the enzymes that catalyze the sulfation of E1, E2 and DHEA and TS and TL PST is unclear. The authors compared thermal stability, sensitivity to inhibition by 2,6-dichloro-4-nitrophenol (DCNP) and individual variation in the regulation of these steroid ST activities with those of TS PST and TL PST in the human liver. E2 ST and TS PST had very similar thermal stabilities. The thermal inactivation profile of E1 ST suggested that this activity might be related to both DHEA ST and TS PST. DCNP inhibition studies also showed similar profiles for E2 ST and TS PST, with a small resistant component for E2 ST. A multiphasic profile for DCNP inhibition of E1 ST activity was found. Finally, studies performed with human liver sample showed significant correlations between E2 ST and TS PST, E1 ST and DHEA ST, E2 St and E1 ST, and, to a lesser degree, between E1 ST and TS PST and E2 ST and DHEA ST. TL PST was not correlated significantly with any of the other activities. These results suggest that the sulfation of E2 in human liver is catalyzed predominantly by TS PST, although DHEA ST may also play a role. Their results also suggest that the sulfation of E1 is catalyzed by DHEA ST and by TS PST, although other ST(s) could also be involved.

  19. A systems biology analysis of the changes in gene expression via silencing of HPV-18 E1 expression in HeLa cells.

    Science.gov (United States)

    Castillo, Andres; Wang, Lu; Koriyama, Chihaya; Eizuru, Yoshito; Jordan, King; Akiba, Suminori

    2014-10-01

    Previous studies have reported the detection of a truncated E1 mRNA generated from HPV-18 in HeLa cells. Although it is unclear whether a truncated E1 protein could function as a replicative helicase for viral replication, it would still retain binding sites for potential interactions with different host cell proteins. Furthermore, in this study, we found evidence in support of expression of full-length HPV-18 E1 mRNA in HeLa cells. To determine whether interactions between E1 and cellular proteins play an important role in cellular processes other than viral replication, genome-wide expression profiles of HPV-18 positive HeLa cells were compared before and after the siRNA knockdown of E1 expression. Differential expression and gene set enrichment analysis uncovered four functionally related sets of genes implicated in host defence mechanisms against viral infection. These included the toll-like receptor, interferon and apoptosis pathways, along with the antiviral interferon-stimulated gene set. In addition, we found that the transcriptional coactivator E1A-binding protein p300 (EP300) was downregulated, which is interesting given that EP300 is thought to be required for the transcription of HPV-18 genes in HeLa cells. The observed changes in gene expression produced via the silencing of HPV-18 E1 expression in HeLa cells indicate that in addition to its well-known role in viral replication, the E1 protein may also play an important role in mitigating the host's ability to defend against viral infection.

  20. The repression domain of the E1B 55-kilodalton protein participates in countering interferon-induced inhibition of adenovirus replication.

    Science.gov (United States)

    Chahal, Jasdave S; Gallagher, Courtney; DeHart, Caroline J; Flint, S J

    2013-04-01

    To begin to investigate the mechanism by which the human adenovirus type 5 E1B 55-kDa protein protects against the antiviral effects of type 1 interferon (IFN) (J. S. Chahal, J. Qi, and S. J. Flint, PLoS Pathog. 8:e1002853, 2012 [doi:10.1371/journal.ppat.1002853]), we examined the effects of precise amino acid substitution in this protein on resistance of viral replication to the cytokine. Only substitution of residues 443 to 448 of E1B for alanine (E1B Sub19) specifically impaired production of progeny virus and resulted in a large defect in viral DNA synthesis in IFN-treated normal human fibroblasts. Untreated or IFN-treated cells infected by this mutant virus (AdEasyE1Sub19) contained much higher steady-state concentrations of IFN-inducible GBP1 and IFIT2 mRNAs than did wild-type-infected cells and of the corresponding newly transcribed pre-mRNAs, isolated exploiting 5'-ethynyluridine labeling and click chemistry. These results indicated that the mutations created by substitution of residues 443 to 448 for alanine (Sub19) impair repression of transcription of IFN-inducible genes, by the E1B, 55-kDa protein, consistent with their location in a segment required for repression of p53-dependent transcription. However, when synthesized alone, the E1B 55-kDa protein inhibited expression of the p53-regulated genes BAX and MDM2 but had no impact whatsoever on induction of IFIT2 and GBP1 expression by IFN. These observations correlate repression of transcription of IFN-inducible genes by the E1B 55-kDa protein with protection against inhibition of viral genome replication and indicate that the E1B 55-kDa protein is not sufficient to establish such transcriptional repression.

  1. 11 CFR 300.52 - Fundraising by Federal candidates and Federal officeholders (2 U.S.C. 441i(e)(1)&(4)).

    Science.gov (United States)

    2010-01-01

    ... 11 Federal Elections 1 2010-01-01 2010-01-01 false Fundraising by Federal candidates and Federal officeholders (2 U.S.C. 441i(e)(1)&(4)). 300.52 Section 300.52 Federal Elections FEDERAL ELECTION COMMISSION... Fundraising by Federal candidates and Federal officeholders (2 U.S.C. 441i(e)(1)&(4)). A Federal candidate,...

  2. Structures of Human Cyctochrome P450 2E1: Insights Into the Binding of Inhibitors And Both Small Molecular Weight And Fatty Acid Substrates

    Energy Technology Data Exchange (ETDEWEB)

    Porubsky, P.R.; Meneely, K.M.; Scott, E.E.

    2009-05-21

    Human microsomal cytochrome P-450 2E1 (CYP2E1) monooxygenates >70 low molecular weight xenobiotic compounds, as well as much larger endogenous fatty acid signaling molecules such as arachidonic acid. In the process, CYP2E1 can generate toxic or carcinogenic compounds, as occurs with acetaminophen overdose, nitrosamines in cigarette smoke, and reactive oxygen species from uncoupled catalysis. Thus, the diverse roles that CYP2E1 has in normal physiology, toxicity, and drug metabolism are related to its ability to metabolize diverse classes of ligands, but the structural basis for this was previously unknown. Structures of human CYP2E1 have been solved to 2.2 {angstrom} for an indazole complex and 2.6 {angstrom} for a 4-methylpyrazole complex. Both inhibitors bind to the heme iron and hydrogen bond to Thr{sup 303} within the active site. Complementing its small molecular weight substrates, the hydrophobic CYP2E1 active site is the smallest yet observed for a human cytochrome P-450. The CYP2E1 active site also has two adjacent voids: one enclosed above the I helix and the other forming a channel to the protein surface. Minor repositioning of the Phe{sup 478} aromatic ring that separates the active site and access channel would allow the carboxylate of fatty acid substrates to interact with conserved {sup 216}QXXNN{sup 220} residues in the access channel while positioning the hydrocarbon terminus in the active site, consistent with experimentally observed {omega}-1 hydroxylation of saturated fatty acids. Thus, these structures provide insights into the ability of CYP2E1 to effectively bind and metabolize both small molecule substrates and fatty acids.

  3. Alcohol dehydrogenase and cytochrome P450 2E1 can be induced by long-term exposure to ethanol in cultured liver HEP-G2 cells.

    Science.gov (United States)

    Balusikova, Kamila; Kovar, Jan

    2013-09-01

    It has been shown in previous studies that liver HEP-G2 cells (human hepatocellular carcinoma) lose their ability to express active alcohol dehydrogenase (ADH) and cytochrome P450 2E1 (CYP2E1). Although both are ethanol-inducible enzymes, short-term exposure to ethanol does not cause any changes in expression or activity in cultured HEP-G2 cells. Therefore, we tested the effect of long-term exposure to ethanol on the expression and activity of both ADH and CYP2E1 in these cells. The expression of ADH and CYP2E1 was assessed at the mRNA and/or protein level using real-time PCR and Western blot analysis. Specific colorimetric assays were used for the measurement of ADH and CYP2E1 enzymatic activities. Caco-2 cells (active CYP2E1 and inactive ADH) were used as control cells. Significantly increased protein expression of ADH (about 2.5-fold) as well as CYP2E1 (about 1.6-fold) was found in HEP-G2 cells after long-term (12 mo) exposure to ethanol. The activity of ADH and CYP2E1 was also significantly increased from 12 ± 3 and 6 ± 1 nmol/h/mg of total protein to 191 ± 9 and 57 ± 9 nmol/h/mg of total protein, respectively. We suggest that the loss of activity of ethanol-metabolizing enzymes in cultured HEP-G2 cells is reversible and can be induced by prolonged exposure to ethanol. We are therefore able to reactivate HEP-G2 cells metabolic functions concerning ethanol oxidation just by modification of in vitro culture conditions without necessity of transfection with its side effect - enzyme overexpression.

  4. Independent regions of adenovirus E1A are required for binding to and dissociation of E2F-protein complexes

    DEFF Research Database (Denmark)

    Fattaey, A R; Harlow, E; Helin, K

    1993-01-01

    overexpression of pRB or p107 in cells lacking a functional pRB leads to the repression of E2F activity. The products of the adenovirus E1A gene can disrupt E2F complexes and result in free and presumably active E2F transcription factor. The regions of E1A required for this function are also essential...

  5. Construction of a mouse Aos1-Uba2 chimeric SUMO-E1 enzyme, mAU, and its expression in baculovirus-insect cells

    Science.gov (United States)

    Nakayama, Tomofumi; Yuasa, Eri; Kanemaru, Ayumi; Saito, Masayuki; Saitoh, Hisato

    2014-01-01

    Small ubiquitin-related modifier (SUMO) is a highly conserved protein that is covalently attached to target proteins. This posttranslational modification, designated SUMOylation, is a major protein-conjugation-driven strategy designed to regulate structure and function of cellular proteins. SUMOylation consists of an enzymatic cascade involving the E1-activating enzyme and the E2-conjugating enzyme. The SUMO-E1 enzyme consists of two subunits, a heterodimer of activation of Smt3p 1 (Aos1) and ubiquitin activating enzyme 2 (Uba2), which resembles the N- and C-terminal halves of ubiquitin E1 (Uba1). Herein, we describe the rational design of a single polypeptide version of SUMO-E1, a chimera of mouse Aos1 and Uba2 subunits, termed mAU, in which the functional domains appear to be arranged in a fashion similar to Uba1. We also describe the construction of a mAU plasmid for expression in a baculovirus-insect cell system and present an in situ SUMOylation assay using the recombinant mAU. Our results showed that mAU has SUMO-E1 activity, thereby indicating that mAU can be expressed in baculovirus-insect cells and represents a suitable source of SUMO-E1. PMID:24637489

  6. The Search For The Cp-violating Emission Of An E1 Photon From The Kaon Long Decays To Positive Pion Negative Pion Gamma

    CERN Document Server

    Shields, J M

    2005-01-01

    A search for the CP-violating electric dipole (E1) direct emission contribution to the KL → π+π −γ decay is performed using data from the 1997 KTeV/E832 experiment. Because the KL → π +π−γ decay mode is massively dominated by the CP-violating inner bremsstrahlung (IB) and the CP-conserving magnetic dipole (M1) direct emission processes, previous analyses have neglected the E1 contribution. Therefore, this measurement is the first attempt to directly quantify the size of the E1 decay process. This E1 transition is one of the very few CP-violating processes that is accessible to experiment and, in principle, will produce new insights into the structure of the neutral kaon. The result of this analysis is that the E1 contribution is below the threshold of sensitivity, and therefore an upper bound of |g E1| < 0.14 (90% CL) is reported. In the process of obtaining this upper limit, high resolution measurements of fit parame...

  7. MECP2e1 isoform mutation affects the form and function of neurons derived from Rett syndrome patient iPS cells.

    Science.gov (United States)

    Djuric, Ugljesa; Cheung, Aaron Y L; Zhang, Wenbo; Mok, Rebecca S; Lai, Wesley; Piekna, Alina; Hendry, Jason A; Ross, P Joel; Pasceri, Peter; Kim, Dae-Sung; Salter, Michael W; Ellis, James

    2015-04-01

    MECP2 mutations cause the X-linked neurodevelopmental disorder Rett Syndrome (RTT) by consistently altering the protein encoded by the MECP2e1 alternative transcript. While mutations that simultaneously affect both MECP2e1 and MECP2e2 isoforms have been widely studied, the consequence of MECP2e1 deficiency on human neurons remains unknown. Here we report the first isoform-specific patient induced pluripotent stem cell (iPSC) model of RTT. RTTe1 patient iPS cell-derived neurons retain an inactive X-chromosome and express only the mutant allele. Single-cell mRNA analysis demonstrated they have a molecular signature of cortical neurons. Mutant neurons exhibited a decrease in soma size, reduced dendritic complexity and decreased cell capacitance, consistent with impaired neuronal maturation. The soma size phenotype was rescued cell-autonomously by MECP2e1 transduction in a level-dependent manner but not by MECP2e2 gene transfer. Importantly, MECP2e1 mutant neurons showed a dysfunction in action potential generation, voltage-gated Na(+) currents, and miniature excitatory synaptic current frequency and amplitude. We conclude that MECP2e1 mutation affects soma size, information encoding properties and synaptic connectivity in human neurons that are defective in RTT.

  8. MC3T3-E1 cell response to stainless steel 316L with different surface treatments

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Hongyu [State Key Laboratory of Tribology, Department of Mechanical Engineering, Tsinghua University, Beijing 100084 (China); Han, Jianmin, E-mail: siyanghan@163.com [Dental Materials Laboratory, National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing 100081 (China); Sun, Yulong [State Key Laboratory of Tribology, Department of Mechanical Engineering, Tsinghua University, Beijing 100084 (China); Huang, Yongling [Jinghang Biomedicine Engineering Division, Beijing Institute of Aeronautical Material, Beijing 100095 (China); Zhou, Ming [State Key Laboratory of Tribology, Department of Mechanical Engineering, Tsinghua University, Beijing 100084 (China)

    2015-11-01

    In the present study, stainless steel 316L samples with polishing, aluminum oxide blasting, and hydroxyapatite (HA) coating were prepared and characterized through a scanning electron microscope (SEM), optical interferometer (surface roughness, Sq), contact angle, surface composition and phase composition analyses. Osteoblast-like MC3T3-E1 cell adhesion on the samples was investigated by cell morphology using a SEM (4 h, 1 d, 3 d, 7 d), and cell proliferation was assessed by MTT method at 1 d, 3 d, and 7 d. In addition, adsorption of bovine serum albumin on the samples was evaluated at 1 h. The polished sample was smooth (Sq: 1.8 nm), and the blasted and HA coated samples were much rougher (Sq: 3.2 μm and 7.8 μm). Within 1 d of incubation, the HA coated samples showed the best cell morphology (e.g., flattened shape and complete spread), but there was no significant difference after 3 d and 7 d of incubation for all the samples. The absorbance value for the HA coated samples was the highest after 1 d and 3 d of incubation, indicating better cell viability. However, it reduced to the lowest value at 7 d. Protein adsorption on the HA coated samples was the highest at 1 h. The results indicate that rough stainless steel surface improves cell adhesion and morphology, and HA coating contributes to superior cell adhesion, but inhibits cell proliferation. - Highlights: • Rough stainless steel surface improves cell adhesion and proliferation. • HA coating results in superior cell morphology and cell attachment. • HA coating inhibits osteoblast cell proliferation after 7 d of incubation.

  9. Rigorous Photogrammetric Processing of CHANG'E-1 and CHANG'E-2 Stereo Imagery for Lunar Topographic Mapping

    Science.gov (United States)

    Di, K.; Liu, Y.; Liu, B.; Peng, M.

    2012-07-01

    Chang'E-1(CE-1) and Chang'E-2(CE-2) are the two lunar orbiters of China's lunar exploration program. Topographic mapping using CE-1 and CE-2 images is of great importance for scientific research as well as for preparation of landing and surface operation of Chang'E-3 lunar rover. In this research, we developed rigorous sensor models of CE-1 and CE-2 CCD cameras based on push-broom imaging principle with interior and exterior orientation parameters. Based on the rigorous sensor model, the 3D coordinate of a ground point in lunar body-fixed (LBF) coordinate system can be calculated by space intersection from the image coordinates of con-jugate points in stereo images, and the image coordinates can be calculated from 3D coordinates by back-projection. Due to uncer-tainties of the orbit and the camera, the back-projected image points are different from the measured points. In order to reduce these inconsistencies and improve precision, we proposed two methods to refine the rigorous sensor model: 1) refining EOPs by correcting the attitude angle bias, 2) refining the interior orientation model by calibration of the relative position of the two linear CCD arrays. Experimental results show that the mean back-projection residuals of CE-1 images are reduced to better than 1/100 pixel by method 1 and the mean back-projection residuals of CE-2 images are reduced from over 20 pixels to 0.02 pixel by method 2. Consequently, high precision DEM (Digital Elevation Model) and DOM (Digital Ortho Map) are automatically generated.

  10. Preliminary Analysis of MicroRNAs Expression Profiling in MC3T3-E1 Cells Exposed to Fluoride.

    Science.gov (United States)

    Wang, Yan; Zhang, Xiuyun; Zhao, Zhitao; Xu, Hui

    2017-04-01

    Overexposure to fluoride from environmental sources can cause serious public health problems. Disrupted osteoblast function and impaired bone formation were found to be associated with excessive fluoride exposure. A massive analysis of microRNAs (miRNAs) was used to figure out the possible pathways in which fluoride affects osteoblast function. MC3T3-E1 cells were treated with 8 mg/L of fluorine for 7 days. Total RNA of cells was extracted, and their integrity and purity were tested. RNA samples were analyzed by using miRNA array, including miRNA labeling, hybridization, scanning, and expression data analysis to compare the profiling of miRNA expression between control and fluoride-treated group. Transcriptome analysis console and enrichment analysis calculated by miRSystem were used to predict target genes and collect miRNAs pathway maps. Forty-five upregulated and 31 downregulated miRNAs expression were found in the fluoride-treated group, and most of the verified miRNAs were mature. The KEGG pathway enrichment analysis searched out 36 pathways that scored more than 0.1. These pathways mainly included intracellular signaling, cytokines, metabolism, and cytoskeleton-related pathways. Among them, the Wnt, insulin, TGF-beta, hedgehog, VEGF, and notch pathways in osteoblasts were those mainly affected by fluoride treatment. These results have shown a number of higher level systemic pathways activated by overexposure of fluoride in osteoblastic cells and verified that fluoride affected the molecular crosstalk in the osteoblasts.

  11. Does prostaglandin-E1 modulate d-galactosamine induced cell death in primary culture of human hepatocytes?

    Directory of Open Access Journals (Sweden)

    Sameh S. Tawfik

    2015-12-01

    Full Text Available Background: Cell death pathway can occur under physiological or pathological conditions. In vitro and in vivo studies have shown that d-galactosamine (DGA induces hepatocyte damage. Objective: The present study aims to evaluate the protective effect of prostaglandin E1 (PGE1 on DGA-induced apoptosis, necrosis and oxidative stress in primary culture of human hepatocytes. Methods: Normal human hepatocytes were obtained from the safety margin of liver specimens, removed during hepatectomy operation to liver cancer patients, and isolated using the classical collagenase perfusion method. After culture stabilization, PGE1 (1 μM was added 2 h before DGA (5 mM. Cultures were maintained for 24 h before the parameters for apoptosis, necrosis and oxidative stress were measured. Apoptosis was studied by DNA-fragmentation, neutral (nSMase and acid (aSMase sphingomyelinase and caspase-3 activity. Necrosis was investigated by lactate dehydrogenase (LDH and transaminases (ALT & AST enzymes. The oxidative stress was assessed by malondialdehyde (MDA, glutathione (GSH, oxidized glutathione (GSSH, glutathione S-transferase (GST, glutathione peroxidase (GSPx, catalase (CAT, superoxide dismutase (SOD and nitric oxide (NO. Results: The hepatotoxin DGA induced apoptosis and enhanced all parameters related to necrosis and intracellular oxidative stress. On the other hand, PGE1 reduced the measured values for the parameters indicative for the DAG induced apoptosis, necrosis and oxidative stress. In addition, PGE1 proved also to prevent GSH depletion. The obtained results provided evidences for the biochemical hepatotoxic effects of DGA (5 mM especially through the induction of apoptosis, necrosis and oxidative stress alterations in the cultured human hepatocytes. Conclusion: PGE1 could be a useful protective treatment against DGA-induced hepatocyte cell death.

  12. Cardioprotective effect of liposomal prostaglandin E1 on a porcine model of myocardial infarction reperfusion no-reflow

    Institute of Scientific and Technical Information of China (English)

    Jia-hui LI; Peng YANG; Ai-li LI; Yong WANG; Yuan-nan KE; Xian-lun LI

    2011-01-01

    Abstract:Objective:To evaluate whether liposomal prostaglandin E1 (lipo-PGE1) can decrease reperfusion no-reflow in a catheter-based porcine model of acute myocardial infarction (AMI).Methods:Twenty-two male Chinese mini-swines were randomized into three groups:six in a sham-operation group,and eight each in the control and lipo-PGE1 groups.The distal part of the left anterior descending coronary artery (LAD) in the latter two groups was completely occluded for 2 h,and then reperfused for 3 h.Lipo-PGE1 (1 pg/kg) was injected 10 min before LAD occlusion until reperfusion for 1 h in the lipo-PGE1 group.Hemodynamic data and proinflammatory cytokines were examined before AMI,2 h after occlusion,and 1,2,and 3 h after reperfusion.Myocardial contrast echocardiography (MCE) and double staining were performed to evaluate the myocardial no-reflow area (NRA).Results:Left ventricular systolic pressure and end-diastolic pressure significantly improved in the lipo-PGE1 group after reperfusion compared with the control group and also 2 h after AMI (P<0.05 for both).MCE and double staining both showed that lipo-PGE1 decreased reperfusion NRA after AMI (P<0.05,P<0.01).Lipo-PGE1 decreased serum interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) after myocardial infarction reperfusion (P<0.05 for both).Conclusions:Lipo-PGE1 is cardioprotective in our porcine model of myocardial infarction reperfusion no-reflow,decreasing NRA and attenuating the inflammatory response.

  13. Reducing error in feline platelet enumeration by addition of Iloprost to blood specimens: comparison to prostaglandin E1 and EDTA.

    Science.gov (United States)

    Tvedten, Harold W; Bäcklund, Kerstin; Lilliehöök, Inger E

    2015-06-01

    Prostaglandin E1 (PGE1) and Iloprost inhibit platelet aggregation and should prevent or minimize preanalytic error with feline platelet enumeration. The objective was to compare the relative effectiveness in reducing errors in platelet enumeration by adding Iloprost to feline EDTA blood specimens in comparison to adding PGE1 or EDTA alone. In addition, a grading system for platelet aggregation in blood smears was evaluated for effectiveness in predicting prominent errors and compared to ADVIA's PLT-CLM flag. Finally, the use of plateletcrit in feline blood with platelet aggregation was evaluated. Blood specimens from 35 cats were included. Blood was collected into EDTA tubes with or without Iloprost or PGE1, and was rapidly mixed. Platelet count (PLT), plateletcrit (PCT), mean platelet volume (MPV), and platelet flags were determined with an ADVIA 2120. Manual PLT was performed with a Leucoplate stain. PLT was determined by an IDEXX VetAutoread hematology analyzer (QBC). Neither addition of Iloprost nor PGE1 to EDTA blood specimens completely prevented platelet aggregation. Iloprost-treated specimens had the least severe aggregation. PGE1 was better than EDTA alone. Significant errors in PLT results were consistently identified by the grading system. ADVIA's PLT-CL flag usually predicted significant errors in PLT. QBC PLT results showed high imprecision. Manual PLT error was smaller than ADVIA PLT in EDTA specimens with aggregation. Adding Iloprost to feline blood specimens improved platelet enumeration accuracy. A grading system for severity of platelet aggregation and usually the ADVIA's PLT-CL alarm predicted specimens with significant errors in platelet enumeration. © 2015 American Society for Veterinary Clinical Pathology.

  14. Effect of Complexation with Hydroxylpropyl-β-Cyclodextrin on Solubility, Dissolution Rate and Chemical Stability of Prostaglandin E1

    Institute of Scientific and Technical Information of China (English)

    GUFu-gen; CUIFu-de; GAOYong-liang

    2004-01-01

    Aim To study the effect of complexation with hydroxylpmpyl-β-cycledextrin (HP-β-CD) on the solubility, dissolution rate and chemical stability of pmstaglandin E1 (PGE1), thereby providing a basis for preparing a stable solid or aqueous preparation of PGF1 formulated with HP-β-CD. Methods The effect of HP-β-CD on the solubility of PGF1 was studied by phase solubility method. The formation of inclusion complexes of PGE1 with HP-β-CD in the aqueous solution was confirmed by UV spectra, circular dichroism spectroscopy, and that in the solid state by IR spectra and X-ray diffractometry. An solid inclusion complex of PGF1 with HP-β-CD was prepared by lyophilization. The dissolution rate and stability of the inclusion complex were determined and compared with those of PGE1 alone. Meanwhile, the stability of PGF1 aqueous solutions in the presence of HP-β-CD was studied under different pH conditions. Results The solubility of PGF1 increased linearly with increasing HP-β-CD concentration in various pH buffered solutions, showing typical AL-type phase solubility diagrams. The stability and dissolution rate of the solid inclusion complex of PGE1 were significantly increased, compared with those of pure PGE1 . The stability of PGEI in HP-β-CD solutions was also obviously improved under acidic and basic conditions, but the stabilizing effect was absent under neutral conditions. Conclulsions The solubility, dissolution rate and chemical stability of PGE1 are markedly improved by complexation with HP-β-CD. It is quite possible to prepare a stable PGE1 inclusion complex-containing solid dosage forms, but almost impossible to obtain a stable aqueous preparation of PGE1 formulated with HP-β-CD.

  15. Protective Effects of N-acetylcysteine and a Prostaglandin E1 Analog, Alprostadil, Against Hepatic Ischemia: Reperfusion Injury in Rats.

    Science.gov (United States)

    Hsieh, Cheng-Chu; Hsieh, Shu-Chen; Chiu, Jen-Hwey; Wu, Ying-Ling

    2014-01-01

    Ischemia-reperfusion (I/R) injury has a complex pathophysiology resulting from a number of contributing factors. Therefore, it is difficult to achieve effective treatment or protection by individually targeting the mediators or mechanisms. Our aim was to analyze the individual and combined effects of N-acetylcysteine (NAC) and the prostaglandin E1 (PGE1) analog alprostadil on hepatic I/R injury in rats. Thirty male Sprague-Dawley rats were randomly divided into five groups (six rats per group) as follows: Control group, I/R group, I/R + NAC group, I/R + alprostadil group, and I/R + NAC + alprostadil group. The rats received injections of NAC (150 mg/kg) and/or alprostadil (0.05 μg/kg) over a period of 30 min prior to ischemia. These rats were then subjected to 60 min of hepatic ischemia followed by a 60-min reperfusion period. Hepatic superoxide dismutase (SOD), catalase, and glutathione levels were significantly decreased as a result of I/R injury, but they were increased in groups treated with NAC. Hepatic malondialdehyde (MDA), myeloperoxidase (MPO), and nitric oxide (NO) activities were significantly increased after I/R injury, but they were decreased in the groups with NAC treatment. Alprostadil decreased NO production, but had no effect on MDA and MPO. Histological results showed that both NAC and alprostadil were effective in improving liver tissue morphology during I/R injury. Although NAC and alprostadil did not have a synergistic effect, our findings suggest that treatment with either NAC or alprostadil has benefits for ameliorating hepatic I/R injury.

  16. Effect of prostaglandins E1, E2, and F2 alpha on osteoclast formation in mouse bone marrow cultures

    Energy Technology Data Exchange (ETDEWEB)

    Collins, D.A.; Chambers, T.J. (St. George' s Hospital Medical School, London (United Kingdom))

    1991-02-01

    Prostaglandins (PG) act as direct inhibitors of mature osteoclasts, but although resorption-inhibition is also observed initially PG increase bone resorption in organ culture. This suggests that PG influence bone resorption in organ culture through actions on cell types other than mature osteoclasts. We have therefore tested the effects of PG E1, E2, and F2 alpha on the differentiation of osteoclastic phenotype in mouse bone marrow cultures using bone resorption and calcitonin receptors (CTR) as markers of osteoclastic differentiation. We found that PGE2 (10{sup {minus} 6}-10{sup {minus} 9} M) and PGE1 (10{sup {minus} 6} - 10{sup {minus} 7} M) induced a significant increase in CTR-positive cell numbers, to levels five to eight times those seen in controls and similar to the number induced by 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). Bone resorption was increased (10{sup {minus} 7} M PGE2 and 10{sup {minus} 6} M PGE1) in association with the increased CTR-positive cell numbers, suggesting that the PG also induced resorptive function. 1,25-(OH)2D3 increased both the number of CTR-positive cells and the extent of resorption per cell; the additional presence of PG did not affect the number of CTR-positive cells but did reduce bone resorption compared with 1,25-(OH)2D3 alone. PGF2 alpha had no significant effect on CTR-positive cell induction or bone resorption. The results suggest that PGE1 and E2 induce osteoclastic differentiation in mouse bone marrow cultures and inhibit the function of the osteoclasts thus formed.

  17. Periodontal repair in dogs: histologic observations of guided tissue regeneration with a prostaglandin E1 analog/methacrylate composite.

    Science.gov (United States)

    Trombelli, L; Lee, M B; Promsudthi, A; Guglielmoni, P G; Wikesjö, U M

    1999-06-01

    This report describes observations of healing following guided tissue regeneration (GTR) including surgical implantation of the prostaglandin E1 analog misoprostol with calcium-layered methacrylate particles. Critical size, supra-alveolar periodontal defects were surgically created around the 3rd and 4th mandibular premolar teeth in 4 beagle dogs. Wound management included soaking with a 24 microg/ml misoprostol solution and implantation of the misoprostol/methacrylate composite. One jaw quadrant per animal was prepared for GTR using expanded polytetrafluoroethylene membranes. The gingival flaps were coronally advanced and sutured to submerge the teeth. The tissues covering the surgical sites daily received topical misoprostol in an oral adhesive over the 4-week healing interval. Upon euthanasia, tissue blocks were prepared for histometric analysis of regeneration of alveolar bone and cementum, root resorption and ankylosis. The defect area underneath the membrane and the density of methacrylate particles were recorded for the GTR defects. The methacrylate particles appeared encapsulated in a dense connective tissue without signs of an inflammatory reaction, some in contact to newly formed bone. Alveolar bone regeneration height averaged (+/-SD) 1.2+/-1.0 and 1.0+/-0.6 mm for GTR and non-GTR defects, respectively. Corresponding values for bone regeneration area were 1.3+/-1.0 and 0.7+/-0.5 mm2. Cementum regeneration was confined to the apical aspect of the defects. Small areas of root resorption and ankylosis were observed for all teeth. Bone regeneration area correlated positively to the defect area and negatively to the density of methacrylate particles in the GTR defects. The histologic observations suggest that the methacrylate composite has marginal potential to promote bone and cementum regeneration under provisions for GTR.

  18. Effectiveness of prostaglandin E1 in patients with mixed arterial and venous ulcers of the lower limbs.

    Science.gov (United States)

    De Caridi, Giovanni; Massara, Mafalda; Stilo, Francesco; Spinelli, Francesco; Grande, Raffaele; Butrico, Lucia; de Franciscis, Stefano; Serra, Raffaele

    2016-10-01

    Mixed arterial and venous ulcers of the lower limbs are present in around 15-30% of patients with chronic venous ulcers (CVUs) and are considered difficult-to-heal wounds. The aim of this study was to evaluate the results of the treatment of mixed arterial and venous ulcers of the lower limbs with prostaglandin E1 (PGE1) infusion. This study was carried out in 48 consecutive patients. Patients who showed intolerability to PGE1, and patients with peripheral neuropathy, blood or systemic diseases, malignancy and acute wound infections or necrotic tissue on the wound bed were excluded. The patients were separated at random into two main groups: group I (25 patients) received standard treatment and PGE1 infusion. Group II (23 patients) received only standard treatment. Pre-treatment data indicated the area of ulceration. The number of healed ulcers and the variation in the area of ulceration were considered as endpoints. The endpoints were noticed after 120 days from the beginning of treatment. Healing occurred in 80% of limbs of group I and in 52·2% of limbs of group II patients. The average reduction in area was 92% versus 60% in patients of group I and II, respectively. During the whole treatment period, the incidence of adverse events was 8% in group I: there was one case of headache and one case of headache and hypotension combined. No side effects were recorded in patients of group II. In conclusion, PGE1 infusion is a determinant in the reduction of the healing time of mixed ulcers of the lower limbs.

  19. Prostaglandin E1 Increases Microcirculation in Random Pattern Flaps on Rats Measured with Laser Doppler Perfusion Imaging

    Science.gov (United States)

    Sneistrup, Christian; Berg, Thomas Moe

    2017-01-01

    Background: Reconstructive procedures with pedicled and free flaps are associated with a risk of ischemia. Prostaglandin E1 (PGE1) is a hormone-like substance with known antiischemic and tissue-protective effects. The aim of this study was to evaluate the effect of PGE1 on the microcirculation in random pattern skin flaps on rats. Methods: Twenty-four rats were divided into 2 groups: an intervention group given PGE1 for 6 hours and a control group given saline. The flap (2 × 10 cm) was created on the back of the rats, and the microcirculation was monitored with laser Doppler perfusion imaging in 5 different zones (1, proximal; 5, distal) before surgery and after 60, 180, and 360 minutes postoperatively. Results: Before surgery, there was no difference in the perfusion in any zones between the intervention group and the control group. The mean perfusion values in zone 1 in the intervention group were significantly higher than those in the control group at 60, 180, and 360 minutes postoperatively (P = 0.02, P = 0.05, and P = 0.04, respectively). At 360 minutes, we also found significantly higher levels of perfusion in the intervention group in zones 4 and 5 (P = 0.05 and P = 0.03, respectively) compared with the controls. Comparing the perfusion at 360 to 60 minutes in the intervention group, we found a significant increase in microcirculation in all zones, which were not seen in the control group. Conclusion: PGE1 increased perfusion in the dermal random pattern flaps on rats. PMID:28203503

  20. Functional PstI/RsaI polymorphism in CYP2E1 is associated with the development, progression and poor outcome of gastric cancer.

    Directory of Open Access Journals (Sweden)

    Jin Feng

    Full Text Available BACKGROUND: Cytochrome P450 2E1 (CYP2E1, an ethanol-inducible enzyme, has been shown to metabolically activate various carcinogens, which is critical for the development and progression of cancers. It has demonstrated that CYP2E1 polymorphisms alter the transcriptional activity of the gene. However, studies on the association between CYP2E1 polymorphisms (PstI/RsaI or DraI and gastric cancer have reported conflicting results. Thus, the aim of the present study was to investigate whether CYP2E1 polymorphisms is associated with the development and progression of gastric cancer and its prognosis in Chinese patients. METHODS: A case-control study was conducted in which CYP2E1 PstI/RsaI and DraI polymorphisms were analyzed in 510 Chinese patients with gastric cancer and 510 age- and sex- matched healthy controls by PCR-RFLP. Odds ratios were estimated by multivariate logistic regression, and the lifetime was calculated by Kaplan-Meier survival curves. In addition, a meta-analysis was also conducted to verify the findings. RESULTS: For CYP2E1 PstI/RsaI polymorphism, C2C2 homozygotes (OR = 2.15; CI: 1.18-3.94 and C2 carriers (OR = 1.48; CI: 1.13-1.96 were associated with an increased risk of gastric cancer when compared with C1C1 homozygotes. Both C1C2 and C2C2 genotypes were associated with advanced stage, but not the grade of gastric cancer. Moreover, C2C2 genotype was identified as an independent marker of poor overall survival for gastric cancer. However, there was not any significant association between CYP2E1 DraI polymorphism and the risk of gastric cancer. In the meta-analysis, pooled data from 13 studies confirmed that the CYP2E1 PstI/RsaI polymorphism was associated with a significantly increased risk of gastric cancer. CONCLUSION: CYP2E1 PstI/RsaI polymorphism is associated with increased risk of development, progression and poor prognosis of gastric cancer in Chinese patients. Pooled data from 13 studies, mainly in Asian countries, are in

  1. A high throughput screening assay to screen for CYP2E1 metabolism and inhibition using a fluorogenic vivid p450 substrate.

    Science.gov (United States)

    Marks, Bryan D; Smith, Ronald W; Braun, Heidi A; Goossens, Tony A; Christenson, Marie; Ozers, Mary S; Lebakken, Connie S; Trubetskoy, Olga V

    2002-11-01

    Large-scale screening of multiple compound libraries and combinatorial libraries for pharmacological activity is one of the novel approaches of the modern drug discovery process. The application of isozyme-specific high-throughput screening (HTS) assays for characterizing the interactions of potential drug candidates with major human drug-metabolizing cytochrome p450 enzymes (p450s) is newly becoming an essential part of this process. Fluorescence-based HTS assays have been successfully employed for in vitro assessment of drug-drug interactions and enzyme inhibition with several p450 isoforms, including CYP3A4, CYP2D6, CYP2C9, and CYP2C19. Here we describe a fluorescence-based HTS assay for detecting drug metabolism and inhibition with human CYP2E1. CYP2E1 plays an important role in the metabolism of several drugs, many solvents, and toxins and therefore has been repeatedly linked to numerous pathologies, including cancer, liver and kidney toxicity, diabetes, and alcoholism. The assay is based on the ability of a drug to compete with the fluorogenic Vivid CYP2E1 Blue Substrate for CYP2E1 metabolism and thus enables rapid screening of lead molecules for their inhibitory potential. We have used this assay to screen a panel of drugs and compounds for their effects on CYP2E1 metabolism and inhibition. Our results demonstrate the assay's usefulness in identifying CYP2E1 substrates and inhibitors and in enabling in-depth characterization of their interactions with the CYP2E1 isozyme. We also present detailed characteristics of the assay, including its dynamic range and Z'-factor values, which indicate that this robust assay is well suited for kinetic and inhibition studies in HTS formats.

  2. Crystal structures of a copper(II and the isotypic nickel(II and palladium(II complexes of the ligand (E-1-[(2,4,6-tribromophenyldiazenyl]naphthalen-2-ol

    Directory of Open Access Journals (Sweden)

    Souheyla Chetioui

    2016-08-01

    Full Text Available In the copper(II complex, bis{(E-1-[(2,4,6-tribromophenyldiazenyl]naphthalen-2-olato}copper(II, [Cu(C16H8Br3N2O2], (I, the metal cation is coordinated by two N atoms and two O atoms from two bidentate (E-1-[(2,4,6-tribromophenyldiazenyl]naphthalen-2-olate ligands, forming a slightly distorted square-planar environment. In one of the ligands, the tribromobenzene ring is inclined to the naphthalene ring system by 37.4 (5°, creating a weak intramolecular Cu...Br interaction [3.134 (2 Å], while in the other ligand, the tribromobenzene ring is inclined to the naphthalene ring system by 72.1 (6°. In the isotypic nickel(II and palladium(II complexes, namely bis{(E-1-[(2,4,6-tribromophenyldiazenyl]naphthalen-2-olato}nickel(II, [Ni(C16H8Br3N2O2], (II, and bis{(E-1-[(2,4,6-tribromophenyldiazenyl]naphthalen-2-olato}palladium(II, [Pd(C16H8Br3N2O2], (III, respectively, the metal atoms are located on centres of inversion, hence the metal coordination spheres have perfect square-planar geometries. The tribromobenzene rings are inclined to the naphthalene ring systems by 80.79 (18° in (II and by 80.8 (3° in (III. In the crystal of (I, molecules are linked by C—H...Br hydrogen bonds, forming chains along [010]. The chains are linked by C—H...π interactions, forming sheets parallel to (011. In the crystals of (II and (III, molecules are linked by C—H...π interactions, forming slabs parallel to (10-1. For the copper(II complex (I, a region of disordered electron density was corrected for using the SQUEEZE routine in PLATON [Spek (2015. Acta Cryst. C71, 9–18]. The formula mass and un