Sample records for element 116 293

  1. Isoreserpine promotes {beta}-catenin degradation via Siah-1 up-regulation in HCT116 colon cancer cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gwak, Jungsug; Song, Taeyun; Song, Jie-Young

    2009-09-25

    Aberrant accumulation of intracellular {beta}-catenin in intestinal epithelial cells is a frequent early event during the development of colon cancer. To identify small molecules that decrease the level of intracellular {beta}-catenin, we performed cell-based chemical screening using genetically engineered HEK293 reporter cells to detect compounds that inhibit TOPFlash reporter activity, which was stimulated by Wnt3a-conditioned medium. We found that isoreserpine promoted the degradation of intracellular {beta}-catenin by up-regulation of Siah-1 in HEK293 and HCT116 colon cancer cells. Moreover, isoreserpine repressed the expression of {beta}-catenin/T-cell factor (TCF)-dependent genes, such as cyclin D1 and c-myc, resulting in the suppression of HCT116 cellmore » proliferation. Our findings suggest that isoreserpine can potentially be used as a chemotherapeutic agent against colon cancer.« less

  2. Reduced migration of MLH1 deficient colon cancer cells depends on SPTAN1.

    PubMed

    Hinrichsen, Inga; Ernst, Benjamin Philipp; Nuber, Franziska; Passmann, Sandra; Schäfer, Dieter; Steinke, Verena; Friedrichs, Nicolaus; Plotz, Guido; Zeuzem, Stefan; Brieger, Angela

    2014-01-24

    Defects in the DNA mismatch repair (MMR) protein MLH1 are frequently observed in sporadic and hereditary colorectal cancers (CRC). Affected tumors generate much less metastatic potential than the MLH1 proficient forms. Although MLH1 has been shown to be not only involved in postreplicative MMR but also in several MMR independent processes like cytoskeletal organization, the connection between MLH1 and metastasis remains unclear. We recently identified non-erythroid spectrin αII (SPTAN1), a scaffolding protein involved in cell adhesion and motility, to interact with MLH1. In the current study, the interaction of MLH1 and SPTAN1 and its potential consequences for CRC metastasis was evaluated. Nine cancer cell lines as well as fresh and paraffin embedded colon cancer tissue from 12 patients were used in gene expression studies of SPTAN1 and MLH1. Co-expression of SPTAN1 and MLH1 was analyzed by siRNA knock down of MLH1 in HeLa, HEK293, MLH1 positive HCT116, SW480 and LoVo cells. Effects on cellular motility were determined in MLH1 deficient HCT116 and MLH1 deficient HEK293T compared to their MLH1 proficient sister cells, respectively. MLH1 deficiency is clearly associated with SPTAN1 reduction. Moreover, siRNA knock down of MLH1 decreased the mRNA level of SPTAN1 in HeLa, HEK293 as well as in MLH1 positive HCT116 cells, which indicates a co-expression of SPTAN1 by MLH1. In addition, cellular motility of MLH1 deficient HCT116 and MLH1 deficient HEK293T cells was impaired compared to the MLH1 proficient sister clones. Consequently, overexpression of SPTAN1 increased migration of MLH1 deficient cells while knock down of SPTAN1 decreased cellular mobility of MLH1 proficient cells, indicating SPTAN1-dependent migration ability. These data suggest that SPTAN1 levels decreased in concordance with MLH1 reduction and impaired cellular mobility in MLH1 deficient colon cancer cells. Therefore, aggressiveness of MLH1-positive CRC might be related to SPTAN1.

  3. Reduced migration of MLH1 deficient colon cancer cells depends on SPTAN1

    PubMed Central

    2014-01-01

    Introduction Defects in the DNA mismatch repair (MMR) protein MLH1 are frequently observed in sporadic and hereditary colorectal cancers (CRC). Affected tumors generate much less metastatic potential than the MLH1 proficient forms. Although MLH1 has been shown to be not only involved in postreplicative MMR but also in several MMR independent processes like cytoskeletal organization, the connection between MLH1 and metastasis remains unclear. We recently identified non-erythroid spectrin αII (SPTAN1), a scaffolding protein involved in cell adhesion and motility, to interact with MLH1. In the current study, the interaction of MLH1 and SPTAN1 and its potential consequences for CRC metastasis was evaluated. Methods Nine cancer cell lines as well as fresh and paraffin embedded colon cancer tissue from 12 patients were used in gene expression studies of SPTAN1 and MLH1. Co-expression of SPTAN1 and MLH1 was analyzed by siRNA knock down of MLH1 in HeLa, HEK293, MLH1 positive HCT116, SW480 and LoVo cells. Effects on cellular motility were determined in MLH1 deficient HCT116 and MLH1 deficient HEK293T compared to their MLH1 proficient sister cells, respectively. Results MLH1 deficiency is clearly associated with SPTAN1 reduction. Moreover, siRNA knock down of MLH1 decreased the mRNA level of SPTAN1 in HeLa, HEK293 as well as in MLH1 positive HCT116 cells, which indicates a co-expression of SPTAN1 by MLH1. In addition, cellular motility of MLH1 deficient HCT116 and MLH1 deficient HEK293T cells was impaired compared to the MLH1 proficient sister clones. Consequently, overexpression of SPTAN1 increased migration of MLH1 deficient cells while knock down of SPTAN1 decreased cellular mobility of MLH1 proficient cells, indicating SPTAN1-dependent migration ability. Conclusions These data suggest that SPTAN1 levels decreased in concordance with MLH1 reduction and impaired cellular mobility in MLH1 deficient colon cancer cells. Therefore, aggressiveness of MLH1-positive CRC might be related to SPTAN1. PMID:24456667

  4. Heat shock proteins HSP70 and MRJ cooperatively regulate cell adhesion and migration through urokinase receptor.

    PubMed

    Lin, Yuli; Peng, Nana; Zhuang, Hongqin; Zhang, Di; Wang, Yao; Hua, Zi-Chun

    2014-08-30

    The urokinase-type plasminogen activator receptor (uPAR) is an important regulator of ECM proteolysis, cell-ECM interactions and cell signaling. uPAR and heat shock proteins HSP70 and MRJ (DNAJB6) have been implicated in tumor growth and metastasis. We have reported recently that MRJ (DNAJB6, a heat shock protein) can interact with uPAR and enhance cell adhesion. Here, we identified another heat shock protein HSP70 as a novel uPAR-interacting protein. We performed co-immunoprecipitation in human embryonic kidney (HEK) 293 and colon cancer HCT116 cells as well as immunofluorence assays in HEK293 cells stably transfected with uPAR to investigate the association of suPAR with HSP70/MRJ. To understand the biological functions of the triple complex of suPAR/HSP70/MRJ, we determined whether HSP70 and/or MRJ regulated uPAR-mediated cell invasion, migration, adhesion to vitronectin and MAPK pathway in two pair of human tumor cells (uPAR negative HEK293 cells vs HEK293 cells stably transfected with uPAR and HCT116 cells stably transfected with antisense-uPAR vs HCT116 mock cells transfected with vector only) using transwell assay, wound healing assay, quantitative RT-PCR analyzing mmp2 and mmp9 transcription levels, cell adhesion assay and Western blotting assay. HSP70 and MRJ formed a triple complex with uPAR and over-expression of MRJ enhanced the interaction between HSP70 and uPAR, while knockdown of MRJ decreased soluble uPAR in HCT116 cells (P < 0.05) and reduced the formation of the triple complex, suggesting that MRJ may act as an uPAR-specific adaptor protein to link uPAR to HSP70. Further experiments showed that knockdown of HSP70 and/or MRJ by siRNA inhibited uPAR-mediated cell adhesion to vitronectin as well as suppressed cell invasion and migration. Knockdown of HSP70 and/or MRJ inhibited expression of invasion related genes mmp2 and mmp9. Finally, HSP70 and/or MRJ up-regulated phosphorylation levels of ERK1/2 and FAK suggesting MAPK pathway was involved. All the biological function experiments in cell level showed an additive effect when HSP70 and MRJ were regulated simultaneously indicating their collaborated regulation effects on uPAR. These findings may offer a novel insight into the interactions between uPAR and HSP70/MRJ and their functions in cell adhesion and migration may provide more understanding of the roles in regulating cancer metastasis.

  5. IND2, a pyrimido[1”,2”:1,5]pyrazolo[3,4-b]quinoline derivative, circumvents multi-drug resistance and causes apoptosis in colon cancer cells

    PubMed Central

    Karthikeyan, Chandrabose; Lee, Crystal; Moore, Joshua; Mittal, Roopali; Suswam, Esther A.; Abbott, Kodye L; Pondugula, Satyanarayana R.; Manne, Upender; Narayanan, Narayanan K.; Trivedi, Piyush; Tiwari, Amit K.

    2014-01-01

    Naturally occurring condensed quinolines have anticancer properties. In efforts to find active analogues, we designed and synthesized eight polycyclic heterocycles with a pyrimido[1”,2”:1,5]pyrazolo[3,4-b]quinoline framework (IND series). The compounds were evaluated for activity against colon (HCT-116 and S1-MI-80), prostate (PC3 and DU-145), breast (MCF-7 and MDAMB-231), ovarian (ov2008 and A2780), and hepatocellular (HepG2) cancer cells and against non-cancerous Madin Darby canine kidney (MDCK), mouse embryonic fibroblast (NIH/3T3), and human embryonic kidney cells (HEK293). IND-2, a 4-chloro-2-methyl pyrimido[1”,2”:1,5]pyrazolo[3,4-b]quinoline, exhibited more than tenfold selectivity and potent cytotoxic activity against colon cancer cells relative to the other cancer and non-cancer cells. With five additional colon cancer cell lines (HT-29, HCT-15, LS-180, LS-174, and LoVo), IND-2 had similar cytotoxicity and selectivity, and submicromolar concentrations caused changes in the morphology of HCT-116 and HCT-15 cells. IND-2 did not activate the transactivating function of the pregnane X receptor (PXR), indicating that it does not induce PXR-regulated ABCB1 or ABCG2 transporters. Indeed, IND-2 was not a substrate of ABCB1 or ABCG2, and it induced cytotoxicity in HEK293 cells overexpressing ABCB1 or ABCG2 to the same extent as in normal HEK293 cells. IND-2 was cytotoxic to resistant colon carcinoma S1-MI-80 cells, approximately three- and fivefold more than SN-38 and topotecan, respectively. In HCT-116 colon cancer cells, IND-2 produced concentration-dependent changes in mitochondrial membrane potential, leading to apoptosis, and sub-micromolar concentrations caused chromosomal DNA fragmentation. These findings suggest that, by increasing apoptosis, IND-2 has potential therapeutic efficacy for colorectal cancer. PMID:25537531

  6. Genome-wide profiling of chemoradiation‑induced changes in alternative splicing in colon cancer cells.

    PubMed

    Xiong, Wei; Gao, Depei; Li, Yunfeng; Liu, Xin; Dai, Peiling; Qin, Jiyong; Wang, Guanshun; Li, Kangming; Bai, Han; Li, Wenhui

    2016-10-01

    Alternative splicing is a key mechanism that regulates protein diversity and has been found to be associated with colon cancer progression and metastasis. However, the function of alternative splicing in chemoradiation‑resistant colon cancer remains elusive. In this study, we constructed a chemoradiation‑resistant colon cancer cell line. Through RNA-sequencing of normal and chemoradiation‑resistant colon cancer cells (HCT116), we found 818 genes that were highly expressed in the normal HCT116 cells, whereas 285 genes were highly expressed in the chemoradiation-resistant HCT116 (RCR-HCT116) cells. Gene ontology (GO) analysis showed that genes that were highly expressed in the HCT116 cells were enriched in GO categories related to cell cycle and cell division, whereas genes that were highly expressed in the RCR-HCT116 cells were associated with regulation of system processes and response to wounding. Analysis of alternative splicing events revealed that exon skipping was significantly increased in the chemoradiation‑resistant colon cancer cells. Moreover, we identified 323 alternative splicing events in 293 genes that were significantly different between the two different HCT116 cell types. These alternative splicing‑related genes were clustered functionally into several groups related with DNA replication, such as deoxyribonucleotide metabolic/catabolic processes, response to DNA damage stimulus and helicase activity. These findings enriched our knowledge by elucidating the function of alternative splicing in chemoradiation-resistant colon cancer.

  7. 32 CFR 11.6 - Crimes and elements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Defense Department of Defense OFFICE OF THE SECRETARY OF DEFENSE MILITARY COMMISSIONS CRIMES AND ELEMENTS FOR TRIALS BY MILITARY COMMISSION § 11.6 Crimes and elements. (a) Substantive offenses—war crimes. The... object of the attack was civilian property, that is, property that was not a military objective; (C) The...

  8. 32 CFR 11.6 - Crimes and elements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Defense Department of Defense OFFICE OF THE SECRETARY OF DEFENSE MILITARY COMMISSIONS CRIMES AND ELEMENTS FOR TRIALS BY MILITARY COMMISSION § 11.6 Crimes and elements. (a) Substantive offenses—war crimes. The... object of the attack was civilian property, that is, property that was not a military objective; (C) The...

  9. 32 CFR 11.6 - Crimes and elements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Defense Department of Defense OFFICE OF THE SECRETARY OF DEFENSE MILITARY COMMISSIONS CRIMES AND ELEMENTS FOR TRIALS BY MILITARY COMMISSION § 11.6 Crimes and elements. (a) Substantive offenses—war crimes. The... object of the attack was civilian property, that is, property that was not a military objective; (C) The...

  10. Short Interspersed Nuclear Element (SINE) Sequences in the Genome of the Human Pathogenic Fungus Aspergillus fumigatus Af293

    PubMed Central

    Kanhayuwa, Lakkhana; Coutts, Robert H. A.

    2016-01-01

    Novel families of short interspersed nuclear element (SINE) sequences in the human pathogenic fungus Aspergillus fumigatus, clinical isolate Af293, were identified and categorised into tRNA-related and 5S rRNA-related SINEs. Eight predicted tRNA-related SINE families originating from different tRNAs, and nominated as AfuSINE2 sequences, contained target site duplications of short direct repeat sequences (4–14 bp) flanking the elements, an extended tRNA-unrelated region and typical features of RNA polymerase III promoter sequences. The elements ranged in size from 140–493 bp and were present in low copy number in the genome and five out of eight were actively transcribed. One putative tRNAArg-derived sequence, AfuSINE2-1a possessed a unique feature of repeated trinucleotide ACT residues at its 3’-terminus. This element was similar in sequence to the I-4_AO element found in A. oryzae and an I-1_AF long nuclear interspersed element-like sequence identified in A. fumigatus Af293. Families of 5S rRNA-related SINE sequences, nominated as AfuSINE3, were also identified and their 5'-5S rRNA-related regions show 50–65% and 60–75% similarity to respectively A. fumigatus 5S rRNAs and SINE3-1_AO found in A. oryzae. A. fumigatus Af293 contains five copies of AfuSINE3 sequences ranging in size from 259–343 bp and two out of five AfuSINE3 sequences were actively transcribed. Investigations on AfuSINE distribution in the fungal genome revealed that the elements are enriched in pericentromeric and subtelomeric regions and inserted within gene-rich regions. We also demonstrated that some, but not all, AfuSINE sequences are targeted by host RNA silencing mechanisms. Finally, we demonstrated that infection of the fungus with mycoviruses had no apparent effects on SINE activity. PMID:27736869

  11. Short Interspersed Nuclear Element (SINE) Sequences in the Genome of the Human Pathogenic Fungus Aspergillus fumigatus Af293.

    PubMed

    Kanhayuwa, Lakkhana; Coutts, Robert H A

    2016-01-01

    Novel families of short interspersed nuclear element (SINE) sequences in the human pathogenic fungus Aspergillus fumigatus, clinical isolate Af293, were identified and categorised into tRNA-related and 5S rRNA-related SINEs. Eight predicted tRNA-related SINE families originating from different tRNAs, and nominated as AfuSINE2 sequences, contained target site duplications of short direct repeat sequences (4-14 bp) flanking the elements, an extended tRNA-unrelated region and typical features of RNA polymerase III promoter sequences. The elements ranged in size from 140-493 bp and were present in low copy number in the genome and five out of eight were actively transcribed. One putative tRNAArg-derived sequence, AfuSINE2-1a possessed a unique feature of repeated trinucleotide ACT residues at its 3'-terminus. This element was similar in sequence to the I-4_AO element found in A. oryzae and an I-1_AF long nuclear interspersed element-like sequence identified in A. fumigatus Af293. Families of 5S rRNA-related SINE sequences, nominated as AfuSINE3, were also identified and their 5'-5S rRNA-related regions show 50-65% and 60-75% similarity to respectively A. fumigatus 5S rRNAs and SINE3-1_AO found in A. oryzae. A. fumigatus Af293 contains five copies of AfuSINE3 sequences ranging in size from 259-343 bp and two out of five AfuSINE3 sequences were actively transcribed. Investigations on AfuSINE distribution in the fungal genome revealed that the elements are enriched in pericentromeric and subtelomeric regions and inserted within gene-rich regions. We also demonstrated that some, but not all, AfuSINE sequences are targeted by host RNA silencing mechanisms. Finally, we demonstrated that infection of the fungus with mycoviruses had no apparent effects on SINE activity.

  12. lH-Pyrazolo[3,4-b]quinolin-3-amine derivatives inhibit growth of colon cancer cells via apoptosis and sub G1 cell cycle arrest.

    PubMed

    Karthikeyan, Chandrabose; Amawi, Haneen; Viana, Arabela Guedes; Sanglard, Leticia; Hussein, Noor; Saddler, Maria; Ashby, Charles R; Moorthy, N S Hari Narayana; Trivedi, Piyush; Tiwari, Amit K

    2018-07-15

    A series of lH-pyrazolo[3,4-b]quinolin-3-amine derivatives were synthesized and evaluated for anticancer efficacy in a panel of ten cancer cell lines, including breast (MDAMB-231 and MCF-7), colon (HCT-116, HCT-15, HT-29 and LOVO), prostate (DU-145 and PC3), brain (LN-229), ovarian (A2780), and human embryonic kidney (HEK293) cells, a non-cancerous cell line. Among the eight derivatives screened, compound QTZ05 had the most potent and selective antitumor efficacy in the four colon cancer cell lines, with IC 50 values ranging from 2.3 to 10.2 µM. Furthermore, QTZ05 inhibited colony formation in HCT-116 cells in a concentration-dependent manner. Cell cycle analysis data indicated that QTZ05 caused an arrest in the sub G1 cell cycle in HCT-116 cells. QTZ05 induced apoptosis in HCT-116 cells in a concentration-dependent manner that was characterized by chromatin condensation and increase in the fluorescence of fluorochrome-conjugated Annexin V. The findings from our study suggest that QTZ05 may be a valuable prototype for the development of chemotherapeutics targeting apoptotic pathways in colorectal cancer cells. Copyright © 2018 Elsevier Ltd. All rights reserved.

  13. Experimental challenges for the measurement of the 116Cd(20Ne,20O)116Sn double charge exchange reaction at 15 AMeV

    NASA Astrophysics Data System (ADS)

    Carbone, D.; Cappuzzello, F.; Agodi, C.; Cavallaro, M.; Acosta, L.; Bonanno, D.; Bongiovanni, D.; Borello, T.; Boztosun, I.; Calabrese, S.; Calvo, D.; Chávez Lomelí, E. R.; Deshmukh, N.; de Faria, P. N.; Finocchiaro, P.; Fisichella, M.; Foti, A.; Gallo, G.; Hacisalihoglu, A.; Iazzi, F.; Introzzi, R.; Lanzalone, G.; Linares, R.; Longhitano, F.; Lo Presti, D.; Medina, N.; Muoio, A.; Oliveira, J. R. B.; Pakou, A.; Pandola, L.; Pinna, F.; Reito, S.; Russo, G.; Santagati, G.; Sgouros, O.; Solakcı, S. O.; Soukeras, V.; Souliotis, G.; Spatafora, A.; Torresi, D.; Tudisco, S.; Yildirim, A.; Zagatto, V. A. B.;

    2018-05-01

    The knowledge of the nuclear matrix elements (NME) entering in the expression of the half-life of the neutrinoless double beta decay is fundamental for neutrino physics. Information on the nuclear matrix elements can be obtained by measuring the absolute cross section of double charge exchange nuclear reactions. The two processes present some similarities, the initial and final-state wave functions are the same and the transition operators are similar. The experimental measurements of double charge exchange reactions induced by heavy ions present a number of challenging aspects, since such reactions are characterized by very low cross sections. Such difficulties are discussed for the measurement of the 116Cd(20Ne,20O)116Sn reaction at 15 AMeV.

  14. Evaluation of the major royal jelly proteins as an alternative to fetal bovine serum in culturing human cell lines.

    PubMed

    Chen, Di; Xin, Xiao-Xuan; Qian, Hao-Cheng; Yu, Zhang-Yin; Shen, Li-Rong

    2016-06-01

    Royal jelly (RJ) is a well-known bioactive substance. It contains large amounts of major royal jelly proteins (MRJPs), which express growth-factor-like activity in several animal and human cell lines. However, the question on whether MRJPs possess growth-factor-like activity on all types of cell cultures remains. In order to determine whether MRJPs can be used as an alternative to fetal bovine serum (FBS) in different types of human cell culture, the proliferation of the complex serum with different ratios of MRJPs/FBS (M/F) was evaluated on five cell lines: 293T, HFL-I, 231, HCT116, and Changliver using MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay. The proliferation activity of the combination of the complex M/F serum with cytokines on the test cell lines was also measured. The results demonstrated that the complex serum with M/F 6/4 possessed the highest proliferation activity similar to or in excess of FBS. However, no activity of complex medium with M/F 6/4 was observed in 231 cells, indicating a selectivity of MRJPs on cell types. Compared with the complex medium with M/F 6/4, the complex medium with M/F 6/4 together with two cytokines, epidermal growth factor (EGF) and insulin-transferrin-selenium (ITS), promoted proliferations of Changliver, 293T, HCT116, and HFL-I by 18.73%‒56.19% (P<0.01). Our findings demonstrate that MRJPs could partially replace FBS in culturing many human cell lines.

  15. Single cell array impedance analysis in a microfluidic device

    NASA Astrophysics Data System (ADS)

    Altinagac, Emre; Taskin, Selen; Kizil, Huseyin

    2016-10-01

    Impedance analysis of single cells is presented in this paper. Following the separation of a target cell type by dielectrophoresis in our previous work, this paper focuses on capturing the cells as a single array and performing impedance analysis to point out the signature difference between each cell type. Lab-on-a-chip devices having a titanium interdigitated electrode layer on a glass substrate and a PDMS microchannel are fabricated to capture each cell in a single form and perform impedance analysis. HCT116 (homosapiens colon colorectal carcin) and HEK293 (human embryonic kidney) cells are used in our experiments.

  16. Measurements of the {sup 116}Cd(p,n) and {sup 116}Sn(n,p) reactions at 300 MeV for studying Gamow-Teller transition strengths in the intermediate nucleus of the {sup 116}Cd double-{beta} decay

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sasano, M.; Kuboki, H.; Sekiguchi, K.

    2009-11-09

    The double differential cross sections for the {sup 116}Cd(p,n) and {sup 116}Sn(n,p) reactions at 300 MeV have been measured over a wide excitation-energy region including Gamow-Teller (GT) giant resonance (GTGR) for studying GT transition strengths in the intermediate nucleus of the {sup 116}Cd double-{beta} decay, namely {sup 116}In. A large amount of the strengths in the {beta}{sup +} direction has been newly found in the energy region up to 30 MeV, which may imply that the GT strengths in the GTGR region contribute to the nuclear matrix element of the two-neutrino double-{beta} decay.

  17. HD 54272, a classical λ Bootis star and γ Doradus pulsator

    NASA Astrophysics Data System (ADS)

    Paunzen, E.; Skarka, M.; Holdsworth, D. L.; Smalley, B.; West, R. G.

    2014-05-01

    We detect the second known λ Bootis star (HD 54272) which exhibits γ Doradus-type pulsations. The star was formerly misidentified as a RR Lyrae variable. The λ Bootis stars are a small group (only 2 per cent) of late B to early F-type, Population I stars which show moderate to extreme (up to a factor 100) surface underabundances of most Fe-peak elements and solar abundances of lighter elements (C, N, O, and S). The photometric data from the Wide Angle Search for Planets (WASP) and All Sky Automated Survey (ASAS) projects were analysed. They have an overlapping time base of 1566 d and 2545 d, respectively. Six statistically significant peaks were identified (f1 = 1.410 116 d-1, f2 = 1.283 986 d-1, f3 = 1.293 210 d-1, f4 = 1.536 662 d-1, f5 = 1.157 22 d-1 and f6 = 0.226 57 d-1). The spacing between f1 and f2, f1 and f4, f5 and f2 is almost identical. Since the daily aliasing is very strong, the interpretation of frequency spectra is somewhat ambiguous. From spectroscopic data, we deduce a high rotational velocity (250 ± 25 km s-1) and a metal deficiency of about -0.8 to -1.1 dex compared to the Sun. A comparison with the similar star, HR 8799, results in analogous pulsational characteristics but widely different astrophysical parameters. Since both are λ Bootis-type stars, the main mechanism of this phenomenon, selective accretion, may severely influence γ Doradus-type pulsations.

  18. 40 CFR Table A-7 to Subpart A of... - Data Elements That Are Inputs to Emission Equations and for Which the Reporting Deadline Is March...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ....116(b) Only annual production by product from each EAF (No CEMS). K 98.116(e)(4) All. K 98.116(e)(5... factor if using Eq. Y-3. Y 98.256(e)(10) Only fraction of carbon in the flare gas contributed by methane... methane in coking gas. Y 98.256(l)(5) Only molar volume conversion factor. Y 98.256(m)(3) Only total...

  19. STS-116 and Expedition 12 Preflight Training, VR Lab Bldg. 9.

    NASA Image and Video Library

    2005-05-06

    JSC2005-E-18147 (6 May 2005) --- Astronauts Sunita L. Williams (left), Expedition 14 flight engineer, and Joan E. Higginbotham, STS-116 mission specialist, use the virtual reality lab at the Johnson Space Center to train for their duties aboard the space shuttle. This type of computer interface, paired with virtual reality training hardware and software, helps to prepare the entire team for dealing with space station elements. Williams will join Expedition 14 in progress and serve as a flight engineer after traveling to the station on space shuttle mission STS-116.

  20. Hydrophobic interaction between the SH2 domain and the kinase domain is required for the activation of Csk.

    PubMed

    Mikkola, Esa T; Gahmberg, Carl G

    2010-06-18

    The protein tyrosine kinase C-terminal Src kinase (Csk) is activated by the engagement of its Src homology (SH) 2 domain. However, the molecular mechanism required for this is not completely understood. The crystal structure of the active Csk indicates that Csk could be activated by contact between the SH2 domain and the beta3-alphaC loop in the N-terminal lobe of the kinase domain. To study the importance of this interaction for the SH2-domain-mediated activation of Csk, we mutated the amino acid residues forming the contacts between the SH2 domain and the beta3-alphaC loop. The mutation of the beta3-alphaC loop Ala228 to glycine and of the SH2 domain Tyr116, Tyr133, Leu138, and Leu149 to alanine resulted in the inability of the SH2 domain ligand to activate Csk. Furthermore, the overexpressed Csk mutants A228G, Y133A/Y116A, L138A, and L149A were unable to efficiently inactivate endogenous Src in human embryonic kidney 293 cells. The results suggest that the SH2-domain-mediated activation of Csk is dependent on the binding of the beta3-alphaC loop Ala228 to the hydrophobic pocket formed by the side chains of Tyr116, Tyr133, Leu138, and Leu149 on the surface of the SH2 domain. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  1. Single and double beta decays in the A=100, A=116 and A=128 triplets of isobars

    NASA Astrophysics Data System (ADS)

    Suhonen, J.; Civitarese, O.

    2014-04-01

    In this paper we analyze the ground-state-to-ground-state two-neutrino double beta (2νββ) decays and single EC and β- decays for the A=100 (100Mo-100Tc-100Ru), A=116 (116Cd-116In-116Sn) and A=128 (128Te-128I-128Xe) triplets of isobars. We use the proton-neutron quasiparticle random-phase approximation (pnQRPA) with realistic G-matrix-derived effective interactions in very large single-particle bases. The purpose is to access the effective value of the axial-vector coupling constant gA in the pnQRPA calculations. We show that the three triplets of isobars represent systems with different characteristics of orbital occupancies and cumulative 2νββ nuclear matrix elements. Our analysis points to a considerably quenched averaged effective value of ≈0.6±0.2 in the pnQRPA calculations.

  2. Atlantic Coast Hindcast, Deepwater, Significant Wave Information

    DTIC Science & Technology

    1981-01-01

    251 91 ISO 110 20 662 4.00 - 4.4V96 117 90 96 34. 439 4.50 - 4.99 76 f2 41 20 35 254 5.00 - 5.41 48 69 20 6 27 170 3.50 5 .19. 34 .9 4 13 122 6.00...3.6 LARES +++ERE)- 5.4- NO.+- OF-+ CASE ISO AZMT Z 2.4: : +__ +,_ + + +++ + +. ++ -- :--All +i7+ STATION I SEASON I AZIUTH(OEfGEES)=3S9.# - 539.9...6 . , i . , 2 0 .9 0: - - O 0.,0 is 61 96 96 27 293 0.50 - 0.99 212 5 260 116 636 1.00 - 9.,9 .o .7 ISO 61 ,, 5A 1.50 - 0.0 3 30 10 .9.00 -0 4 , . 7

  3. 40 CFR Table A-7 to Subpart A of... - Data Elements That Are Inputs to Emission Equations and for Which the Reporting Deadline Is March...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... mass balance equation. K 98.116(b) Only annual production by product from each EAF (No CEMS). K 98.116... carbon content values, molecular weights for gaseous feedstocks, molecular weights for gaseous products... of gas flared, average molecular weight, carbon content of the flare, and molar volume conversion...

  4. Alternative reverse genetics system for influenza viruses based on a synthesized swine 45S rRNA promoter.

    PubMed

    Wang, Kai; Huang, Qi; Yang, Zhiwei; Qi, Kezong; Liu, Hongmei; Chen, Hongjun

    2017-08-01

    We generated an alternative reverse genetics (RG) system based on a synthesized swine 45S rRNA promoter to rescue the H3N2 subtype swine influenza virus. All eight flanking segment cassettes of A/swine/Henan/7/2010 (H3N2) were amplified with ambisense expression elements from RG plasmids. All segments were then recombined with the pHC2014 vector, which contained the synthesized swine 45S rRNA promoter (spol1) and its terminal sequence (t1) in a pcDNA3 backbone. As a result, we obtained a set of RG plasmids carrying the corresponding eight-segment cassettes. We efficiently generated the H3N2 virus after transfection into 293T/PK15, PK15, and 293T cells. The efficiency of spol1-driven influenza virus rescue in PK15 cells was similar to that in 293T cells by titration using the human pol1 RG system. Our approach suggests that an alternative spol1-based RG system can produce influenza viruses.

  5. 32 CFR 11.6 - Crimes and elements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... must be a direct result of the substance's effect or effects on the human body (e.g., asphyxiation...) The accused caused serious injury to the body or health of one or more persons; (B) The accused... of the body, and serious damage to internal organs. (13) Mutilation or maiming—(i) Elements. (A) The...

  6. 32 CFR 11.6 - Crimes and elements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... must be a direct result of the substance's effect or effects on the human body (e.g., asphyxiation...) The accused caused serious injury to the body or health of one or more persons; (B) The accused... of the body, and serious damage to internal organs. (13) Mutilation or maiming—(i) Elements. (A) The...

  7. Fusion-fission and quasifission of superheavy systems with Z =110 -116 formed in 48Ca-induced reactions

    NASA Astrophysics Data System (ADS)

    Kozulin, E. M.; Knyazheva, G. N.; Itkis, I. M.; Itkis, M. G.; Bogachev, A. A.; Chernysheva, E. V.; Krupa, L.; Hanappe, F.; Dorvaux, O.; Stuttgé, L.; Trzaska, W. H.; Schmitt, C.; Chubarian, G.

    2014-11-01

    Background: In heavy-ion-induced reactions the mechanism leading to the formation of the compound nucleus and the role of quasifission is still not clear. Purpose: Investigation of the quasifission process of superheavy composite systems with Z =110 -116 and comparison with properties of fusion-fission and quasifission of lighter composite systems. Method: Mass and energy distributions of fissionlike fragments formed in the reactions 48Ca+232Th, 238U , 244Pu , and 248Cm at energies near the Coulomb barrier have been measured using the double-arm time-of-flight spectrometer CORSET at the U-400 cyclotron of the FLNR JINR. Results: The most probable fragment masses as well as total kinetic energies and their dispersions in dependence on the interaction energies and ion-target combinations have been studied for asymmetric and symmetric fragments formed in the reactions. The capture cross sections were obtained for the reactions 48Ca+244Pu and 248Cm . The lower limits for fission barriers of 283 -286Cn , 289 -292Fl , and 293 -296Lv compound nuclei were estimated. Conclusions: Analysis of the properties of symmetric fragments has shown that a significant part of these fragments may be attributed to fusion-fission process for the reactions 48Ca +238U , 244Pu , and 248Cm .

  8. Whole-Genome-Sequencing characterization of bloodstream infection-causing hypervirulent Klebsiella pneumoniae of capsular serotype K2 and ST374.

    PubMed

    Wang, Xiaoli; Xie, Yingzhou; Li, Gang; Liu, Jialin; Li, Xiaobin; Tian, Lijun; Sun, Jingyong; Ou, Hong-Yu; Qu, Hongping

    2018-01-01

    Hypervirulent K. pneumoniae variants (hvKP) have been increasingly reported worldwide, causing metastasis of severe infections such as liver abscesses and bacteremia. The capsular serotype K2 hvKP strains show diverse multi-locus sequence types (MLSTs), but with limited genetics and virulence information. In this study, we report a hypermucoviscous K. pneumoniae strain, RJF293, isolated from a human bloodstream sample in a Chinese hospital. It caused a metastatic infection and fatal septic shock in a critical patient. The microbiological features and genetic background were investigated with multiple approaches. The Strain RJF293 was determined to be multilocis sequence type (ST) 374 and serotype K2, displayed a median lethal dose (LD50) of 1.5 × 10 2 CFU in BALB/c mice and was as virulent as the ST23 K1 serotype hvKP strain NTUH-K2044 in a mouse lethality assay. Whole genome sequencing revealed that the RJF293 genome codes for 32 putative virulence factors and exhibits a unique presence/absence pattern in comparison to the other 105 completely sequenced K. pneumoniae genomes. Whole genome SNP-based phylogenetic analysis revealed that strain RJF293 formed a single clade, distant from those containing either ST66 or ST86 hvKP. Compared to the other sequenced hvKP chromosomes, RJF293 contains several strain-variable regions, including one prophage, one ICEKp1 family integrative and conjugative element and six large genomic islands. The sequencing of the first complete genome of an ST374 K2 hvKP clinical strain should reinforce our understanding of the epidemiology and virulence mechanisms of this bloodstream infection-causing hvKP with clinical significance.

  9. Whole-Genome-Sequencing characterization of bloodstream infection-causing hypervirulent Klebsiella pneumoniae of capsular serotype K2 and ST374

    PubMed Central

    Wang, Xiaoli; Xie, Yingzhou; Li, Gang; Liu, Jialin; Li, Xiaobin; Tian, Lijun; Sun, Jingyong; Qu, Hongping

    2018-01-01

    ABSTRACT Hypervirulent K. pneumoniae variants (hvKP) have been increasingly reported worldwide, causing metastasis of severe infections such as liver abscesses and bacteremia. The capsular serotype K2 hvKP strains show diverse multi-locus sequence types (MLSTs), but with limited genetics and virulence information. In this study, we report a hypermucoviscous K. pneumoniae strain, RJF293, isolated from a human bloodstream sample in a Chinese hospital. It caused a metastatic infection and fatal septic shock in a critical patient. The microbiological features and genetic background were investigated with multiple approaches. The Strain RJF293 was determined to be multilocis sequence type (ST) 374 and serotype K2, displayed a median lethal dose (LD50) of 1.5 × 102 CFU in BALB/c mice and was as virulent as the ST23 K1 serotype hvKP strain NTUH-K2044 in a mouse lethality assay. Whole genome sequencing revealed that the RJF293 genome codes for 32 putative virulence factors and exhibits a unique presence/absence pattern in comparison to the other 105 completely sequenced K. pneumoniae genomes. Whole genome SNP-based phylogenetic analysis revealed that strain RJF293 formed a single clade, distant from those containing either ST66 or ST86 hvKP. Compared to the other sequenced hvKP chromosomes, RJF293 contains several strain-variable regions, including one prophage, one ICEKp1 family integrative and conjugative element and six large genomic islands. The sequencing of the first complete genome of an ST374 K2 hvKP clinical strain should reinforce our understanding of the epidemiology and virulence mechanisms of this bloodstream infection-causing hvKP with clinical significance. PMID:29338592

  10. KSC-06pd2287

    NASA Image and Video Library

    2006-10-12

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, the external tank is transferred from the checkout cell for attaching to its twin solid rocket boosters on the mobile launch platform in highbay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6-feet wide and 154-feet tall. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6-feet wide and 154-feet tall. STS-116 will be mission no. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  11. 40 CFR Table A-7 to Subpart A of... - Data Elements That Are Inputs to Emission Equations and for Which the Reporting Deadline Is March...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... mass balance equation. K 98.116(b) Only annual production by product from each EAF (No CEMS). K 98.116... contributed by methane. Y 98.256(f)(7) Only molar volume conversion factor. Y 98.256(f)(10) Only coke burn-off... methane in coking gas. Y 98.256(l)(5) Only molar volume conversion factor. Y 98.256(m)(3) Only total...

  12. Minnesota Water Allocation Law. Volume 1. Commentary on and Abstracts of Treaties, Legislation and Rules.

    DTIC Science & Technology

    1987-09-01

    If applicable) 8c. ADDRESS(City, State, and ZIP Code) 10. SOURCE OF FUNDING NUMBERS ELEMENT NO. NO. NO ACCESSION NO .T 11 TITLE (Include Security...Indians 1-13 Domestic Water Supply on Reservations 1-14 Ground water sources 1-16 Surface water sources 1-16 Commentary on Indian Water Resources 1-17...General’s Office - St. Paul, Minnesota 4 DATA COLLECTION AND REPORT ORGANIZATION Data collected for these volumes result from three primary sources

  13. Manufacturing Technology Program Information System: Functional Description.

    DTIC Science & Technology

    1983-02-01

    D-A27 293 MANUACTURING TECNOLOGY PROGRAM INFORMATIONSSEM: /; FUNCTIONAL DESCRIPTIONU ALOGSTS CsMANAGEMENT NST WASHINGTON DC K WRIGHT ET AL FEB 83...greater use of computer technology in all elements of manu- facturing. - Assure that more effective industrial innovation is stimulated by reducing the...data base. Data communications capabilities (terminals, communications hardware and software) will make it possible for remote users throughout the MT

  14. Ice particles trapped by temperature gradients at mbar pressure

    NASA Astrophysics Data System (ADS)

    Kelling, Thorben; Wurm, Gerhard; Dürmann, Christoph

    2011-11-01

    In laboratory experiments we observe that ice particles (⩽100 μm) entrained in a low pressure atmosphere (˜1 mbar) get trapped by temperature gradients between three reservoirs at different temperature. Confining elements are a peltier element at 250 K (bottom), a liquid nitrogen reservoir at 77 K (top), and the surrounding vacuum chamber at 293 K. Particle levitation and trapping is modeled by an interplay of thermophoresis, photophoresis, and gravity. A number of ice particles are trapped simultaneously in close spatial distance to each other at least up to minutes and are accessible for further experiments.

  15. KSC-06pd2292

    NASA Image and Video Library

    2006-10-13

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, the external tank is mated to its twin solid rocket boosters on the mobile launch platform in high bay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6 feet wide and 154 feet tall. STS-116 will be mission No. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  16. KSC-06pd2291

    NASA Image and Video Library

    2006-10-13

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, the external tank is mated to its twin solid rocket boosters on the mobile launch platform in high bay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6 feet wide and 154 feet tall. STS-116 will be mission No. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  17. KSC-06pd2289

    NASA Image and Video Library

    2006-10-13

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, the external tank is mated to its twin solid rocket boosters on the mobile launch platform in high bay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6-feet wide and 154-feet tall. STS-116 will be mission No. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  18. KSC-06pd2290

    NASA Image and Video Library

    2006-10-13

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, the external tank is mated to its twin solid rocket boosters on the mobile launch platform in high bay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the space shuttle system at 27.6 feet wide and 154 feet tall. STS-116 will be mission No. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  19. KSC-06pd2293

    NASA Image and Video Library

    2006-10-13

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, the external tank is mated to its twin solid rocket boosters on the mobile launch platform in high bay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6 feet wide and 154 feet tall. STS-116 will be mission No. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  20. Finite element validation of stress intensity factor calculation models for thru-thickness and thumb-nail cracks in double edge notch specimens

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Beres, W.; Koul, A.K.

    1994-09-01

    Stress intensity factors for thru-thickness and thumb-nail cracks in the double edge notch specimens, containing two different notch radius (R) to specimen width (W) ratios (R/W = 1/8 and 1/16), are calculated through finite element analysis. The finite element results are compared with predictions based on existing empirical models for SIF calculations. The effects of a change in R/W ratio on SIF of thru-thickness and thumb-nail cracks are also discussed. 34 refs.

  1. Quantifying Stellar Mass Loss with High Angular Resolution Imaging

    DTIC Science & Technology

    2009-02-19

    material – via massive winds, planetary nebulae and supernova explosions – seeding the interstellar medium with heavier elements. Subsequent...of Planetary Nebulae (Harpaz, ApJ, 498,293, (1998)), impacts the pre-explosion characteristic of SNII (Taylor, “The Stars”, Cambridge (1994)), and...A 464, 119) or may have an important role, such as Be Stars, W-R stars, and planetary nebulae . The Future of Interferometric O/IR Imaging. The

  2. Ice particles trapped by temperature gradients at mbar pressure.

    PubMed

    Kelling, Thorben; Wurm, Gerhard; Dürmann, Christoph

    2011-11-01

    In laboratory experiments we observe that ice particles (≤100 μm) entrained in a low pressure atmosphere (~1 mbar) get trapped by temperature gradients between three reservoirs at different temperature. Confining elements are a peltier element at 250 K (bottom), a liquid nitrogen reservoir at 77 K (top), and the surrounding vacuum chamber at 293 K. Particle levitation and trapping is modeled by an interplay of thermophoresis, photophoresis, and gravity. A number of ice particles are trapped simultaneously in close spatial distance to each other at least up to minutes and are accessible for further experiments. © 2011 American Institute of Physics

  3. Influence of Group IV and V Alloying Elements on the Microstructure Engineering and Deformation Behavior in Tantalum Carbides

    DTIC Science & Technology

    2015-04-06

    materials is a mix of covalent, metallic and ionic resulting in high hardness as well as good thermal and electrical conductivity [4-7]. The materials...difficulty in capturing the out-of-plane sigma bonds. Both these deficiencies are well known. Table A.1: Properties of the pure elements Property Ti Zr ...Hf V Nb Ta Graphite Diamond a (Å) 2.93 3.23 3.19 2.98 3.33 3.31 2.47 3.57 c (Å) 4.62 5.18 5.05 7.72 Ecoh (eV) 5.31 6.33 6.48 5.37 6.93 8.20

  4. Studies on the synthesis of isotopes of superheavy element Lv (Z = 116)

    NASA Astrophysics Data System (ADS)

    Santhosh, K. P.; Safoora, V.

    2017-11-01

    The probable projectile-target combinations for the synthesis of superheavy nucleus 296Lv found in the cold valley of 296Lv have been identified by studying the interaction barrier of the colliding nuclei, probability of compound nucleus formation, P_{CN}, and survival probability W_{sur}. At energies near and above the Coulomb barrier, the capture, fusion and evaporation residue (ER) cross sections for the probable combinations for the hot and cold fusion reactions are systematically investigated. By considering intensities of the projectile beams, availabilities of the targets and half lives of the colliding nuclei, the combination 48Ca+248Cm is found to be the most probable projectile-target pair for the synthesis of 296Lv. The calculated maximum value of 2n, 3n, 4n and 5n channel cross section for the reaction 48Ca+248Cm are 0.599 pb, 5.957 pb, 4.805 pb, and 0.065 pb, respectively. Moreover, the production cross sections for the synthesis of isotopes 291-295,298Lv using 48Ca projectile on 243-247,250Cm targets are calculated. Among these reactions, the reactions 48Ca+247Cm → 295Lv and 48Ca+250Cm → 298Lv have maximum production cross section in 3n (10.697 pb) and 4n (12.006 pb) channel, respectively. Our studies on the SHE Lv using the combinations 48Ca+245Cm → 293Lv and 48Ca+248Cm → 296Lv are compared with available experimental data and with other theoretical studies. Our studies are in agreement with experimental data and we hope that these studies will be a guide for the future experiments to synthesize the isotopes of Lv.

  5. Functional analysis of the sea urchin-derived arylsulfatase (Ars)-element in mammalian cells.

    PubMed

    Watanabe, Satoshi; Watanabe, Sachiko; Sakamoto, Naoaki; Sato, Masahiro; Akasaka, Koji

    2006-09-01

    An insulator is a DNA sequence that has both enhancer-blocking activity, through its ability to modify the influence of neighboring cis-acting elements, and a barrier function that protects a transgene from being silenced by surrounding chromatin. Previously, we isolated and characterized a 582-bp-long element from the sea urchin arylsulfatase gene (Ars). This Ars-element was effective in sea urchin and Drosophila embryos and in plant cells. To investigate Ars-element activity in mammalian cells, we placed the element between the cytomegalovirus enhancer and a luciferase (luc) expression cassette. In contrast to controls lacking the Ars-element, NIH3T3 and 293T cells transfected with the element-containing construct displayed reduced luciferase activities. The Ars-element therefore acts as an enhancer-blocking element in mammalian cells. We assessed the barrier activity of the Ars-element using vectors in which a luc expression cassette was placed between two elements. Transfection experiments demonstrated that luc activity in these vectors was approximately ten-fold higher than in vectors lacking elements. Luc activities were well maintained even after 12 weeks in culture. Our observations demonstrate that the Ars-element has also a barrier activity. These results indicated that the Ars-element act as an insulator in mammalian cells.

  6. Recruitment by the Repressor Freud-1 of Histone Deacetylase-Brg1 Chromatin Remodeling Complexes to Strengthen HTR1A Gene Repression.

    PubMed

    Souslova, Tatiana; Mirédin, Kim; Millar, Anne M; Albert, Paul R

    2017-12-01

    Five-prime repressor element under dual repression binding protein-1 (Freud-1)/CC2D1A is genetically linked to intellectual disability and implicated in neuronal development. Freud-1 represses the serotonin-1A (5-HT1A) receptor gene HTR1A by histone deacetylase (HDAC)-dependent or HDAC-independent mechanisms in 5-HT1A-negative (e.g., HEK-293) or 5-HT1A-expressing cells (SK-N-SH), respectively. To identify the underlying mechanisms, Freud-1-associated proteins were affinity-purified from HEK-293 nuclear extracts and members of the Brg1/SMARCCA chromatin remodeling and Sin3A-HDAC corepressor complexes were identified. Pull-down assays using recombinant proteins showed that Freud-1 interacts directly with the Brg1 carboxyl-terminal domain; interaction with Brg1 required the carboxyl-terminal of Freud-1. Freud-1 complexes in HEK-293 and SK-N-SH cells differed, with low levels of BAF170/SMARCC2 and BAF57/SMARCE1 in HEK-293 cells and low-undetectable BAF155/SMARCC1, Sin3A, and HDAC1/2 in SK-N-SH cells. Similarly, by quantitative chromatin immunoprecipitation, Brg1-BAF170/57 and Sin3A-HDAC complexes were observed at the HTR1A promoter in HEK-293 cells, whereas in SK-N-SH cells, Sin3A-HDAC proteins were not detected. Quantifying 5-HT1A receptor mRNA levels in cells treated with siRNA to Freud-1, Brg1, or both RNAs addressed the functional role of the Freud-1-Brg1 complex. In HEK-293 cells, 5-HT1A receptor mRNA levels were increased only when both Freud-1 and Brg1 were depleted, but in SK-N-SH cells, depletion of either protein upregulated 5-HT1A receptor RNA. Thus, recruitment by Freud-1 of Brg1, BAF155, and Sin3A-HDAC complexes appears to strengthen repression of the HTR1A gene to prevent its expression inappropriate cell types, while recruitment of the Brg1-BAF170/57 complex is permissive to 5-HT1A receptor expression. Alterations in Freud-1-Brg1 interactions in mutants associated with intellectual disability could impair gene repression leading to altered neuronal development.

  7. Recruitment by the Repressor Freud-1 of Histone Deacetylase-Brg1 Chromatin Remodeling Complexes to Strengthen HTR1A Gene Repression

    PubMed Central

    Souslova, Tatiana; Mirédin, Kim; Millar, Anne M.

    2017-01-01

    Five-prime repressor element under dual repression binding protein-1 (Freud-1)/CC2D1A is genetically linked to intellectual disability and implicated in neuronal development. Freud-1 represses the serotonin-1A (5-HT1A) receptor gene HTR1A by histone deacetylase (HDAC)-dependent or HDAC-independent mechanisms in 5-HT1A-negative (e.g., HEK-293) or 5-HT1A-expressing cells (SK-N-SH), respectively. To identify the underlying mechanisms, Freud-1-associated proteins were affinity-purified from HEK-293 nuclear extracts and members of the Brg1/SMARCCA chromatin remodeling and Sin3A-HDAC corepressor complexes were identified. Pull-down assays using recombinant proteins showed that Freud-1 interacts directly with the Brg1 carboxyl-terminal domain; interaction with Brg1 required the carboxyl-terminal of Freud-1. Freud-1 complexes in HEK-293 and SK-N-SH cells differed, with low levels of BAF170/SMARCC2 and BAF57/SMARCE1 in HEK-293 cells and low-undetectable BAF155/SMARCC1, Sin3A, and HDAC1/2 in SK-N-SH cells. Similarly, by quantitative chromatin immuno-precipitation, Brg1-BAF170/57 and Sin3A-HDAC complexes were observed at the HTR1A promoter in HEK-293 cells, whereas in SK-N-SH cells, Sin3A-HDAC proteins were not detected. Quantifying 5-HT1A receptor mRNA levels in cells treated with siRNA to Freud-1, Brg1, or both RNAs addressed the functional role of the Freud-1-Brg1 complex. In HEK-293 cells, 5-HT1A receptor mRNA levels were increased only when both Freud-1 and Brg1 were depleted, but in SK-N-SH cells, depletion of either protein upregulated 5-HT1A receptor RNA. Thus, recruitment by Freud-1 of Brg1, BAF155, and Sin3A-HDAC complexes appears to strengthen repression of the HTR1A gene to prevent its expression inappropriate cell types, while recruitment of the Brg1-BAF170/57 complex is permissive to 5-HT1A receptor expression. Alterations in Freud-1-Brg1 interactions in mutants associated with intellectual disability could impair gene repression leading to altered neuronal development. PMID:27914010

  8. Mass Spectral Detection of Diethoxyphospho-Tyrosine Adducts on Proteins from HEK293 Cells Using Monoclonal Antibody depY for Enrichment

    PubMed Central

    2018-01-01

    Chronic illness from exposure to organophosphorus toxicants is hypothesized to involve modification of unknown proteins. Tyrosine in proteins that have no active site serine readily reacts with organophosphorus toxicants. We developed a monoclonal antibody, depY, that specifically recognizes diethoxyphospho-tyrosine in proteins and peptides, independent of the surrounding amino acid sequence. Our goal in the current study was to identify diethoxyphosphorylated proteins in human HEK293 cell lysate treated with chlorpyrifos oxon. Cell lysates treated with chlorpyrifos oxon were recognized by depY antibody in ELISA and capillary electrophoresis based Western blot. Tryptic peptides were analyzed by liquid chromatography tandem mass spectrometry. Liquid chromatography tandem mass spectrometry identified 116 diethoxyphospho-tyrosine peptides from 73 proteins in immunopurified samples, but found only 15 diethoxyphospho-tyrosine peptides from 12 proteins when the same sample was not immunopurified on depY. The most abundant proteins in the cell lysate, histone H4, heat shock 70 kDa protein 1A/1B, heat shock protein HSP 90 β, and α-enolase, were represented by several diethoxyphospho-tyrosine peptides. It was concluded that use of immobilized depY improved the number of diethoxyphospho-tyrosine peptides identified in a complex mixture. The mass spectrometry results confirmed the specificity of depY for diethoxyphospho-tyrosine peptides independent of the context of the modified tyrosine, which means depY could be used to analyze modified proteins in any species. Use of the depY antibody could lead to an understanding of chronic illness from organophosphorus pesticide exposure. PMID:29775289

  9. KSC-06pd2284

    NASA Image and Video Library

    2006-10-12

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, workers prepare to transfer the external tank from the checkout cell for attaching to its twin solid rocket boosters on the mobile launch platform in highbay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6-feet wide and 154-feet tall. STS-116 will be mission no. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  10. KSC-06pd2286

    NASA Image and Video Library

    2006-10-12

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, the external tank is transferred from the checkout cell for attaching to its twin solid rocket boosters on the mobile launch platform in highbay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6-feet wide and 154-feet tall. STS-116 will be mission no. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  11. KSC-06pd2283

    NASA Image and Video Library

    2006-10-12

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, workers prepare to transfer the external tank from the checkout cell for attaching to its twin solid rocket boosters on the mobile launch platform in highbay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6-feet wide and 154-feet tall. STS-116 will be mission no. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  12. KSC-06pd2288

    NASA Image and Video Library

    2006-10-12

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, the external tank is transferred from the checkout cell for attaching to its twin solid rocket boosters on the mobile launch platform in highbay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6-feet wide and 154-feet tall. STS-116 will be mission no. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  13. KSC-06pd2285

    NASA Image and Video Library

    2006-10-12

    KENNEDY SPACE CENTER, FLA. - Inside the Vehicle Assembly Building, workers prepare to transfer the external tank from the checkout cell for attaching to its twin solid rocket boosters on the mobile launch platform in highbay 3 for mission STS-116. The gigantic, rust-colored external tank is the largest element of the Space Shuttle system at 27.6-feet wide and 154-feet tall. STS-116 will be mission no. 20 to the International Space Station and construction flight 12A.1. The mission payload is the SPACEHAB module, the P5 integrated truss structure and other key components. Launch is scheduled for no earlier than Dec. 7. Photo credit: NASA/Jack Pfaller

  14. [Construction of a general AAV vector regulated by minimal and artificial hypoxic-responsive element].

    PubMed

    Nie, Xiao-wei; Sun, Li-jun; Hao, Yue-wen; Yang, Guang-xiao; Wang, Quan-ying

    2011-03-01

    To synthesize the minimal and artificial HRE, and to insert it into the anterior extremity of CMV promoter of a AAV plasmid, and then to construct the AAV regulated by hypoxic-responsive element which was introduced into 293 cell by method of Ca3(PO4)2 using three plasmids. Thus obtaining the adenoassociated virus vector regulated by hypoxic-responsive element was possibly used for gene therapy in ischemia angiocardiopathy and cerebrovascular disease. Artificially synthesize the 36 bp nucleotide sequences of four connection in series HIF-binding sites A/GCGTG(4×HBS)and a 35 bp nucleotide sequences spacing inserted into anterior extremity of CMV promoter TATA Box, then amplified by PCR. The cDNA fragment was confirmed to be right by DNA sequencing. Molecular biology routine method was used to construct a AAV vector regulated by minimal hypoxic-responsive element after the normal CMV promoter in AAV vector was replaced by the CMV promoter included minimal hypoxic-responsive element. Then, NT4-6His-PR39 fusogenic peptide was inserted into MCS of the plasmid, the recombinant AAV vector was obtained by three plasmid co-transfection in 293 cells, in which we can also investigate the expression of 6×His using immunochemistry in hypoxia environment. Artificial HRE was inserted into anterior extremity of CMV promoter and there was a correct spacing between the HRE and the TATA-box. The DNA sequencing and restriction enzyme digestion results indicated that the AAV regulated by hypoxic-responsive element was successfully constructed. Compared to the control group, the expressions of 6×His was significantly increased in the experimental groups in hypoxia environment, which confirmed that the AAV effectually regulated by the minimal HRE was inserted into anterior extremity of CMV promoter. The HRE is inserted into anterior extremity of CMV promoter to lack incision enzyme recognition site by PCR. And eukaryotic expression vector regulated by hypoxic-responsive is constructed. The AAV effectually regulated by the minimal HRE inserted into anterior extremity of CMV promoter. The vector is successfully constructed and it has important theoretical and practical value in the synteresis and therapy of ischemia angiocardiopathy and cerebrovascular disease.

  15. Development of freight policy analysis tool for northeastern Illinois and the United States.

    DOT National Transportation Integrated Search

    2010-10-01

    Freight transportation is a vital element in the economic prosperity of any country. According to the : nationwide commodity flow survey, over 12 billion tons of goods, valued at more than $11.6 trillion, were moved in : America in the year 2007(Bure...

  16. Development and fabrication of an advanced liquid cooling garment

    NASA Technical Reports Server (NTRS)

    Leith, J. R.; Hixon, C. W.

    1976-01-01

    The elastomeric film fin/tube concept which was developed is a composite of polyurethane film, fine expanded silver mesh, a serpentine pattern polyurethane transport tubing and an integral comfort liner, all bonded via adhesive application and vacuum-bagged for final cure. As demonstrated by thermal analysis, the composite garment material is capable of removing a 293 watt (1000 BTU/hr) metabolic load through a head and torso cooling area of .46 sq m (5 sq ft) with tube spacing of slightly under one inch. A total of 60 test elements, each .15m x .15m (6 in. x 6 in.) were fabricated in support of the liquid cooling garment concept development. In parallel with the fabrication of these elements a continuing series of laboratory tests to support the fabrication techniques was carried out. The elements and supporting tests are described.

  17. 32 CFR 219.116 - General requirements for informed consent.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... will not adversely affect the rights and welfare of the subjects; (3) The research could not... subject in research covered by this policy unless the investigator has obtained the legally effective... investigator, the sponsor, the institution or its agents from liability for negligence. (a) Basic elements of...

  18. 14 CFR § 1230.116 - General requirements for informed consent.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... will not adversely affect the rights and welfare of the subjects; (3) The research could not... research covered by this policy unless the investigator has obtained the legally effective informed consent..., the institution or its agents from liability for negligence. (a) Basic elements of informed consent...

  19. U. S. Naval Forces, Vietnam Monthly Historical Summary for February 1970

    DTIC Science & Technology

    1970-04-09

    Search Turn Campaign .................... 28 Riverine Strike Grouo ..... .............. . . . . . . 33 Breezy Cove. ......... . . . . . . . . . 34...116 MVAYAL ADIVYSORY GROUP SUMRY . .. .. .. .. .. .. . .. 119 Vietnamese Mdarine Corps .. .. .. .. .. .. .. ... . 137 VNMC Operations Control ...helicopter (Dustoff) and artillery assistance wias requested. At 1 9377, -an Air Force rors-ard Air Controller arrived on station and air elements

  20. Butyrate induces apoptosis by activating PDC and inhibiting complex I through SIRT3 inactivation.

    PubMed

    Xu, Sha; Liu, Cai-Xia; Xu, Wei; Huang, Lei; Zhao, Jian-Yuan; Zhao, Shi-Min

    2017-01-01

    The underlying anticancer effects of butyrate, an end-product of the intestinal microbial fermentation of dietary fiber, remain elusive. Here, we report that butyrate promotes cancer cell apoptosis by acting as a SIRT3 inhibitor. Butyrate inhibits SIRT3 both in cultured cells and in vitro . Butyrate-induced PDHA1 hyperacetylation relieves the inhibitory phosphorylation of PDHA1 at serine 293, thereby activating an influx of glycolytic intermediates into the tricarboxylic acid (TCA) cycle and reversing the Warburg effect. Meanwhile, butyrate-induced hyperacetylation inactivates complex I of the electron transfer chain and prevents the utilization of TCA cycle intermediates. These metabolic stresses promote apoptosis in hyperglycolytic cancer cells, such as HCT116 p53 -/- cells. SIRT3 deacetylates both PDHA1 and complex I. Genetic ablation of Sirt3 in mouse hepatocytes abrogated the ability of butyrate to induce apoptosis. Our results identify a butyrate-mediated anti-tumor mechanism and indicate that the combined activation of PDC and inhibition of complex I is a novel tumor treatment strategy.

  1. Growth and anisotropic thermal properties of biaxial Ho:YAlO3 crystal

    NASA Astrophysics Data System (ADS)

    Dong, Qin; Zhao, Guangjun; Chen, Jianyu; Ding, Yuchong; Zhao, Chengchun

    2010-07-01

    Ho:YAlO3 (YAP) crystal with large size and good optical quality has been grown by the Czochralski method. Thermal properties of the as-grown Ho:YAP crystal have been investigated by measuring the temperature-dependent, anisotropic thermal expansion, specific heat, thermal diffusion, and thermal conductivity. The results show that Ho:YAP crystal possesses a large anisotropic thermal expansion and good thermal conductivity. The calculated average thermal expansion coefficients along a, b, and c axis are αa=9.18×10-6/K, αb=1.94×10-6/K, and αc=7.61×10-6/K from 293.15 to 770.15 K. The thermal conductivities along a, b, and c axis are up to 11.6, 9.9, and 12.3 W m-1 K-1 at 298.15 K. Compared with Ho: Y3Al5O12 (YAG), Ho:YAP crystal has a larger thermal conductivity along a axis from 298.15 to 568.15 K.

  2. Boehmenan, a lignan from Hibiscus ficulneus, showed Wnt signal inhibitory activity.

    PubMed

    Shono, Takumi; Ishikawa, Naoki; Toume, Kazufumi; Arai, Midori A; Ahmed, Firoj; Sadhu, Samir K; Ishibashi, Masami

    2015-07-15

    The Wnt signal pathway modulates numerous biological processes, and its aberrant activation is related to various diseases. Therefore, inhibition of the Wnt signal may provide an effective (or efficient) strategy for these diseases. Cell-based luciferase assay targeting the Wnt signal (TOP assay) revealed that Hibiscus ficulneus extract inhibited the Wnt signal. The activity-guided isolation of the MeOH extract of H. ficulneus stems yielded four known (1-4) lignans along with myriceric acid (5). Compounds 1-4 potently inhibited the Wnt signal with TOPflash IC50 values of 1.0, 4.5, 6.3, and 1.9 μM, respectively. Compound 1 exhibited cytotoxicity against both Wnt-dependent (HCT116) and Wnt-independent (RKO) cells. Western blot analysis showed that 1 decreased the expression of full, cytosolic and nuclear β-catenin along with c-myc in STF/293 cells. Our results suggested that 1 may have inhibited the Wnt signal by decreasing β-catenin levels. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Hyperforin activates gene transcription involving transient receptor potential C6 channels.

    PubMed

    Thiel, Gerald; Rössler, Oliver G

    2017-04-01

    Hypericum perforatum is one of the most prominent medical plants. Hyperforin, a main ingredient of H. perforatum, has been shown to activate transient receptor potential canonical C6 (TRPC6) channels. Alternatively, it has been proposed that hyperforin functions as a protonophore in a TRPC6-independent manner. Here, we show that hyperforin stimulation activates the transcription factor AP-1 in HEK293 cells expressing TRPC6 (T6.11 cells), but did not substantially change the AP-1 activity in HEK293 cells lacking TRPC6. We identified the AP-1 binding site as a hyperforin-responsive element. AP-1 is composed of the transcription factors c-Jun and c-Fos, or other members of the c-Jun and c-Fos families of proteins. Hyperforin stimulation increased c-Jun and c-Fos promoter activities in T6.11 cells and induced an upregulation of c-Jun and c-Fos biosynthesis. The analysis of the c-Fos promoter revealed that the cAMP-response element also functions as a hyperforin-responsive element. Hyperforin-induced upregulation of AP-1 in T6.11 cells was attenuated by preincubation of the cells with either pregnenolone or progesterone, indicating that gene regulation via TRPC6 is under control of hormones or hormonal precursors. The signal transduction of hyperforin-induced AP-1 gene transcription required an influx of Ca 2+ ions into the cells, the activation of MAP kinases, and the activation of the transcription factors c-Jun and ternary complex factor. We conclude that hyperforin regulates gene transcription via activation of TRPC6 channels, involving stimulus-regulated protein kinases and stimulus-responsive transcription factors. The fact that hyperforin regulates gene transcription may explain many of the intracellular alterations induced by this compound. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. 8 CFR 293.1 - Computation of interest.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... simple interest table in § 293.3 shall be utilized in the computation of interest under this part. ... 8 Aliens and Nationality 1 2010-01-01 2010-01-01 false Computation of interest. 293.1 Section 293... INTEREST ON CASH RECEIVED TO SECURE IMMIGRATION BONDS § 293.1 Computation of interest. Interest shall be...

  5. Genome dynamics of the human embryonic kidney 293 lineage in response to cell biology manipulations.

    PubMed

    Lin, Yao-Cheng; Boone, Morgane; Meuris, Leander; Lemmens, Irma; Van Roy, Nadine; Soete, Arne; Reumers, Joke; Moisse, Matthieu; Plaisance, Stéphane; Drmanac, Radoje; Chen, Jason; Speleman, Frank; Lambrechts, Diether; Van de Peer, Yves; Tavernier, Jan; Callewaert, Nico

    2014-09-03

    The HEK293 human cell lineage is widely used in cell biology and biotechnology. Here we use whole-genome resequencing of six 293 cell lines to study the dynamics of this aneuploid genome in response to the manipulations used to generate common 293 cell derivatives, such as transformation and stable clone generation (293T); suspension growth adaptation (293S); and cytotoxic lectin selection (293SG). Remarkably, we observe that copy number alteration detection could identify the genomic region that enabled cell survival under selective conditions (i.c. ricin selection). Furthermore, we present methods to detect human/vector genome breakpoints and a user-friendly visualization tool for the 293 genome data. We also establish that the genome structure composition is in steady state for most of these cell lines when standard cell culturing conditions are used. This resource enables novel and more informed studies with 293 cells, and we will distribute the sequenced cell lines to this effect.

  6. Glutamine synthetase gene knockout-human embryonic kidney 293E cells for stable production of monoclonal antibodies.

    PubMed

    Yu, Da Young; Lee, Sang Yoon; Lee, Gyun Min

    2018-05-01

    Previously, it was inferred that a high glutamine synthetase (GS) activity in human embryonic kidney (HEK) 293E cells results in elevated resistance to methionine sulfoximine (MSX) and consequently hampers GS-mediated gene amplification and selection by MSX. To overcome this MSX resistance in HEK293E cells, a GS-knockout HEK293E cell line was generated using the CRISPR/Cas9 system to target the endogenous human GS gene. The GS-knockout in the HEK293E cell line (RK8) was confirmed by Western blot analysis of GS and by observation of glutamine-dependent growth. Unlike the wild type HEK293E cells, the RK8 cells were successfully used as host cells to generate a recombinant HEK293E cell line (rHEK293E) producing a monoclonal antibody (mAb). When the RK8 cells were transfected with the GS expression vector containing the mAb gene, rHEK293E cells producing the mAb could be selected in the absence as well as in the presence of MSX. The gene copies and mRNA expression levels of the mAb in rHEK293E cells were also quantified using qRT-PCR. Taken together, the GS-knockout HEK293E cell line can be used as host cells to generate stable rHEK293E cells producing a mAb through GS-mediated gene selection in the absence as well as in the presence of MSX. © 2018 Wiley Periodicals, Inc.

  7. Activation of ERK and JNK signaling pathways by mycotoxin citrinin in human cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chang, C.-H.; Yu, F.-Y.; Wang, L.-T.

    2009-06-15

    Mycotoxin citrinin (CTN) is commonly found in foods and feeds that are contaminated/inoculated with Penicillium, Aspergillus and Monascus species. The exposure of human embryonic kidney (HEK293) and HeLa cells to CTN resulted in a dose-dependent increase in the phosphorylation of two major mitogen-activated protein kinases (MAPKs), ERK1/2 and JNK. In HEK293 cultures, the administering of CTN increased both the mRNA and protein levels of egr-1, c-fos and c-jun genes; additionally, the ERK1/2 pathway contributed to the upregulation of Egr-1 and c-Fos protein expression. CTN treatment also induced the transcription activity of Egr-1 and AP-1 proteins, as evidenced by luciferase reportermore » assays. Bioinformatic analyses indicated two genes Gadd45{beta} and MMP3 have Egr-1 and AP-1 response elements in their promoters, respectively. Furthermore, co-exposure of HEK293 cells to CTN and MAPK pathway inhibitors demonstrated that CTN increased the levels of Gadd45{beta} mRNA through ERK1/2 signaling pathway and up-regulated the MMP3 transcripts majorly via JNK pathway. Finally, CTN-triggered caspase 3 activity was significantly reduced in the presence of MAPK inhibitors. Our results suggest that CTN positively regulates ERK1/2 and JNK pathways as well as their downstream effectors in human cells; activated MAPK pathways are also involved in CTN-induced apoptosis.« less

  8. Medicinal herbs Oenanthe javanica (Blume) DC., Casuarina equisetifolia L. and Sorghum bicolor (L.) Moench protect human cells from MPP+ damage via inducing FBXO7 expression.

    PubMed

    Chen, Chiung-Mei; Chen, I-Cheng; Chen, Ying-Lin; Lin, Te-Hsien; Chen, Wan-Ling; Chao, Chih-Ying; Wu, Yih-Ru; Lu, Yeah-Ting; Lee, Cheng-Yu; Chien, Hong-Chi; Chen, Ting-Shou; Lee-Chen, Guey-Jen; Lee, Chi-Mei

    2016-11-15

    The F-box protein 7 (FBXO7) mutations have been identified in families with early-onset parkinsonism and pyramidal tract signs, and designated as PARK15. In addition, FBXO7 mutations were found in typical and young onset Parkinson's disease (PD). Evidence has also shown that FBXO7 plays an important role in the development of dopaminergic neurons and increased stability and overexpression of FBXO7 may be beneficial to PD. We screened extracts of medicinal herbs to enhance FBXO7 expression for neuroprotection in MPP + -treated cells. Promoter reporter assay in HEK-293 cells was used to examine the cis/trans elements controlling FBXO7 expression and to screen extracts of medicinal herbs enhancing FBXO7 expression. MTT assay was performed to assess cell viability of MPP + -treated HEK-293/SH-SY5Y cells. In addition, proteasome activity, mitochondrial membrane potential and FBXO7/TRAF2/GATA2 protein expression were evaluated. We demonstrated that -202--57 region of the FBXO7 promoter is likely to contain sequences that are bound by positive trans protein factors to activate FBXO7 expression and GATA2 is the main trans protein factor enhancing FBXO7 expression. Extracts of medicinal herbs Oenanthe javanica (Blume) DC. (Umbelliferae), Casuarina equisetifolia L. (Casuarinaceae), and Sorghum bicolor (L.) Moench (Gramineae) improved cell viability of both MPP + -treated HEK-293 and SH-SY5Y cells, rescued proteasome activity in MPP + -treated HEK-293 cells, and restored mitochondrial membrane potential in MPP + -treated SH-SY5Y cells. These protection effects of herbal extracts are acting through enhancing FBXO7 and decreasing TRAF2 expression, which is probably mediated by GATA2 induction. Collectively, our study provides new targets, FBXO7 and its regulator GATA2, for the development of potential treatments of PD. Copyright © 2016 Elsevier GmbH. All rights reserved.

  9. 5 CFR 293.401 - Applicability of regulations.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Applicability of regulations. 293.401 Section 293.401 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.401 Applicability of regulations. This...

  10. 5 CFR 293.401 - Applicability of regulations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Applicability of regulations. 293.401 Section 293.401 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.401 Applicability of regulations. This...

  11. 5 CFR 293.501 - Applicability of regulations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Applicability of regulations. 293.501 Section 293.501 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.501 Applicability of regulations. The...

  12. 5 CFR 293.501 - Applicability of regulations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Applicability of regulations. 293.501 Section 293.501 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.501 Applicability of regulations. The...

  13. 5 CFR 293.401 - Applicability of regulations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Applicability of regulations. 293.401 Section 293.401 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.401 Applicability of regulations. This...

  14. 5 CFR 293.501 - Applicability of regulations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Applicability of regulations. 293.501 Section 293.501 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.501 Applicability of regulations. The...

  15. 5 CFR 293.501 - Applicability of regulations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Applicability of regulations. 293.501 Section 293.501 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.501 Applicability of regulations. The...

  16. 5 CFR 293.401 - Applicability of regulations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Applicability of regulations. 293.401 Section 293.401 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.401 Applicability of regulations. This...

  17. 5 CFR 293.501 - Applicability of regulations.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Applicability of regulations. 293.501 Section 293.501 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.501 Applicability of regulations. The...

  18. 5 CFR 293.401 - Applicability of regulations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Applicability of regulations. 293.401 Section 293.401 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.401 Applicability of regulations. This...

  19. 5 CFR 293.301 - Applicability of regulations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Applicability of regulations. 293.301 Section 293.301 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.301 Applicability of regulations. This subpart applies to...

  20. 5 CFR 293.301 - Applicability of regulations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Applicability of regulations. 293.301 Section 293.301 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.301 Applicability of regulations. This subpart applies to...

  1. 5 CFR 293.106 - Safeguarding information about individuals.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Safeguarding information about individuals. 293.106 Section 293.106 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.106 Safeguarding...

  2. 5 CFR 293.104 - Collection of information.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Collection of information. 293.104 Section 293.104 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.104 Collection of information. (a...

  3. 5 CFR 293.511 - Retention schedule.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Retention schedule. 293.511 Section 293.511 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.511 Retention schedule. (a) Temporary EMFS records must...

  4. 5 CFR 293.511 - Retention schedule.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Retention schedule. 293.511 Section 293.511 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.511 Retention schedule. (a) Temporary EMFS records must...

  5. 5 CFR 293.511 - Retention schedule.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Retention schedule. 293.511 Section 293.511 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.511 Retention schedule. (a) Temporary EMFS records must...

  6. 5 CFR 293.511 - Retention schedule.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Retention schedule. 293.511 Section 293.511 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.511 Retention schedule. (a) Temporary EMFS records must...

  7. 5 CFR 293.511 - Retention schedule.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Retention schedule. 293.511 Section 293.511 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.511 Retention schedule. (a) Temporary EMFS records must...

  8. 28 CFR 115.293 - Audits of standards.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 28 Judicial Administration 2 2012-07-01 2012-07-01 false Audits of standards. 115.293 Section 115.293 Judicial Administration DEPARTMENT OF JUSTICE (CONTINUED) PRISON RAPE ELIMINATION ACT NATIONAL STANDARDS Standards for Community Confinement Facilities Audits § 115.293 Audits of standards. The agency...

  9. 28 CFR 115.293 - Audits of standards.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 2 2013-07-01 2013-07-01 false Audits of standards. 115.293 Section 115.293 Judicial Administration DEPARTMENT OF JUSTICE (CONTINUED) PRISON RAPE ELIMINATION ACT NATIONAL STANDARDS Standards for Community Confinement Facilities Audits § 115.293 Audits of standards. The agency...

  10. 28 CFR 115.293 - Audits of standards.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 28 Judicial Administration 2 2014-07-01 2014-07-01 false Audits of standards. 115.293 Section 115.293 Judicial Administration DEPARTMENT OF JUSTICE (CONTINUED) PRISON RAPE ELIMINATION ACT NATIONAL STANDARDS Standards for Community Confinement Facilities Audits § 115.293 Audits of standards. The agency...

  11. 5 CFR 293.302 - Establishment of Official Personnel Folder.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Establishment of Official Personnel Folder. 293.302 Section 293.302 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.302 Establishment of Official Personnel Folder...

  12. 5 CFR 293.302 - Establishment of Official Personnel Folder.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Establishment of Official Personnel Folder. 293.302 Section 293.302 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.302 Establishment of Official Personnel Folder...

  13. 5 CFR 293.107 - Special safeguards for automated records.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Special safeguards for automated records. 293.107 Section 293.107 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.107 Special safeguards...

  14. 5 CFR 293.510 - Disposition of Employee Medical Folders.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Disposition of Employee Medical Folders. 293.510 Section 293.510 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.510 Disposition of Employee Medical...

  15. 5 CFR 293.510 - Disposition of Employee Medical Folders.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Disposition of Employee Medical Folders. 293.510 Section 293.510 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.510 Disposition of Employee Medical...

  16. 5 CFR 293.102 - Definitions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... not limited to, his or her education, financial transactions, medical history, criminal history, or... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Definitions. 293.102 Section 293.102... Policies on Maintenance of Personnel Records § 293.102 Definitions. In this part: Agency means any...

  17. 5 CFR 293.402 - Establishment of separate employee performance record system.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... performance record system. 293.402 Section 293.402 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.402 Establishment of separate employee performance record system. (a) Copies of employees' performance ratings of...

  18. 5 CFR 293.402 - Establishment of separate employee performance record system.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... performance record system. 293.402 Section 293.402 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.402 Establishment of separate employee performance record system. (a) Copies of employees' performance ratings of...

  19. 5 CFR 293.510 - Disposition of Employee Medical Folders.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Disposition of Employee Medical Folders. 293.510 Section 293.510 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.510 Disposition of Employee Medical...

  20. 5 CFR 293.402 - Establishment of separate employee performance record system.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... performance record system. 293.402 Section 293.402 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.402 Establishment of separate employee performance record system. (a) Copies of employees' performance ratings of...

  1. 5 CFR 293.510 - Disposition of Employee Medical Folders.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Disposition of Employee Medical Folders. 293.510 Section 293.510 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.510 Disposition of Employee Medical...

  2. 5 CFR 293.402 - Establishment of separate employee performance record system.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... performance record system. 293.402 Section 293.402 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.402 Establishment of separate employee performance record system. (a) Copies of employees' performance ratings of...

  3. 5 CFR 293.402 - Establishment of separate employee performance record system.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... performance record system. 293.402 Section 293.402 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.402 Establishment of separate employee performance record system. (a) Copies of employees' performance ratings of...

  4. 5 CFR 293.510 - Disposition of Employee Medical Folders.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Disposition of Employee Medical Folders. 293.510 Section 293.510 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.510 Disposition of Employee Medical...

  5. STS116-S-001

    NASA Image and Video Library

    2006-07-01

    STS116-S-001 (July 2006) --- The STS-116 patch design signifies the continuing assembly of the International Space Station (ISS). The primary mission objective is to deliver and install the P5 truss element. The P5 installation will be conducted during the first of three planned spacewalks, and will involve use of both the shuttle and station robotic arms. The remainder of the mission will include a major reconfiguration and activation of the ISS electrical and thermal control systems, as well as delivery of Zvezda Service Module debris panels, which will increase ISS protection from potential impacts of micro-meteorites and orbital debris. In addition, a single expedition crewmember will launch on STS-116 to remain onboard the station, replacing an expedition crew member that will fly home with the shuttle crew. The crew patch depicts the space shuttle rising above the Earth and ISS. The United States and Swedish flags trail the orbiter, depicting the international composition of the STS-116 crew. The seven stars of the constellation Ursa Major are used to provide direction to the North Star, which is superimposed over the installation location of the P5 truss on ISS. The NASA insignia design for space shuttle flights is reserved for use by the astronauts and for other official use as the NASA Administrator may authorize. Public availability has been approved only in the forms of illustrations by the various news media. When and if there is any change in this policy, which is not anticipated, the change will be publicly announced. Photo credit: NASA

  6. Modulation of voltage- and Ca2+-dependent gating of CaV1.3 L-type calcium channels by alternative splicing of a C-terminal regulatory domain.

    PubMed

    Singh, Anamika; Gebhart, Mathias; Fritsch, Reinhard; Sinnegger-Brauns, Martina J; Poggiani, Chiara; Hoda, Jean-Charles; Engel, Jutta; Romanin, Christoph; Striessnig, Jörg; Koschak, Alexandra

    2008-07-25

    Low voltage activation of Ca(V)1.3 L-type Ca(2+) channels controls excitability in sensory cells and central neurons as well as sinoatrial node pacemaking. Ca(V)1.3-mediated pacemaking determines neuronal vulnerability of dopaminergic striatal neurons affected in Parkinson disease. We have previously found that in Ca(V)1.4 L-type Ca(2+) channels, activation, voltage, and calcium-dependent inactivation are controlled by an intrinsic distal C-terminal modulator. Because alternative splicing in the Ca(V)1.3 alpha1 subunit C terminus gives rise to a long (Ca(V)1.3(42)) and a short form (Ca(V)1.3(42A)), we investigated if a C-terminal modulatory mechanism also controls Ca(V)1.3 gating. The biophysical properties of both splice variants were compared after heterologous expression together with beta3 and alpha2delta1 subunits in HEK-293 cells. Activation of calcium current through Ca(V)1.3(42A) channels was more pronounced at negative voltages, and inactivation was faster because of enhanced calcium-dependent inactivation. By investigating several Ca(V)1.3 channel truncations, we restricted the modulator activity to the last 116 amino acids of the C terminus. The resulting Ca(V)1.3(DeltaC116) channels showed gating properties similar to Ca(V)1.3(42A) that were reverted by co-expression of the corresponding C-terminal peptide C(116). Fluorescence resonance energy transfer experiments confirmed an intramolecular protein interaction in the C terminus of Ca(V)1.3 channels that also modulates calmodulin binding. These experiments revealed a novel mechanism of channel modulation enabling cells to tightly control Ca(V)1.3 channel activity by alternative splicing. The absence of the C-terminal modulator in short splice forms facilitates Ca(V)1.3 channel activation at lower voltages expected to favor Ca(V)1.3 activity at threshold voltages as required for modulation of neuronal firing behavior and sinoatrial node pacemaking.

  7. Mechanism of estrogen activation of c-myc oncogene expression.

    PubMed

    Dubik, D; Shiu, R P

    1992-08-01

    The estrogen receptor complex is a known trans-acting factor that regulates transcription of specific genes through an interaction with a specific estrogen-responsive cis-acting element (ERE). In previous studies we have shown that in estrogen-responsive human breast cancer cells estrogen rapidly activates c-myc expression. This activated expression occurs through enhanced transcription and does not require the synthesis of new protein intermediates; therefore, an ERE is present in the human c-myc gene regulatory region. To localize the ERE, constructs containing varying lengths of the c-myc 5'-flanking region ranging from -2327 to +25 (relative to the P1 promoter) placed adjacent to the chloramphenicol acetyl transferase reporter gene (CAT) were prepared. They were used in transient transfection studies in MCF-7 and HeLa cells co-transfected with an estrogen receptor expression vector. These studies reveal that all constructs containing the P2 promoter region exhibited estrogen-regulated CAT expression and that a 116-bp region upstream and encompassing the P2 TATA box is necessary for this activity. Analysis of this 116-bp region failed to identify a cis-acting element with sequences resembling the consensus ERE; however, co-transfection studies with mutant estrogen receptor expression vectors showed that the DNA-binding domain of the receptor is essential for estrogen-regulated CAT gene expression. We have also observed that anti-estrogen receptor complexes can weakly trans-activate from this 116-bp region but fail to do so from the ERE-containing ApoVLDLII-CAT construct. To explain these results we propose a new mechanism of estrogen trans-activation in the c-myc gene promoter.

  8. 47 CFR 73.293 - Use of FM multiplex subcarriers.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 47 Telecommunication 4 2011-10-01 2011-10-01 false Use of FM multiplex subcarriers. 73.293 Section 73.293 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) BROADCAST RADIO SERVICES RADIO BROADCAST SERVICES FM Broadcast Stations § 73.293 Use of FM multiplex subcarriers. Licensees of FM broadcast...

  9. 47 CFR 73.293 - Use of FM multiplex subcarriers.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 4 2010-10-01 2010-10-01 false Use of FM multiplex subcarriers. 73.293 Section 73.293 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) BROADCAST RADIO SERVICES RADIO BROADCAST SERVICES FM Broadcast Stations § 73.293 Use of FM multiplex subcarriers. Licensees of FM broadcast...

  10. 5 CFR 293.505 - Establishment and protection of Employee Medical Folder.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Establishment and protection of Employee Medical Folder. 293.505 Section 293.505 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.505 Establishment and...

  11. 5 CFR 293.505 - Establishment and protection of Employee Medical Folder.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Establishment and protection of Employee Medical Folder. 293.505 Section 293.505 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.505 Establishment and...

  12. 5 CFR 293.505 - Establishment and protection of Employee Medical Folder.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Establishment and protection of Employee Medical Folder. 293.505 Section 293.505 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.505 Establishment and...

  13. 5 CFR 293.505 - Establishment and protection of Employee Medical Folder.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Establishment and protection of Employee Medical Folder. 293.505 Section 293.505 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.505 Establishment and...

  14. 5 CFR 293.505 - Establishment and protection of Employee Medical Folder.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Establishment and protection of Employee Medical Folder. 293.505 Section 293.505 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.505 Establishment and...

  15. 32 CFR 293.2 - Policy.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 2 2010-07-01 2010-07-01 false Policy. 293.2 Section 293.2 National Defense... PROGRAM NATIONAL IMAGERY MAPPING AGENCY (NIMA) FREEDOM OF INFORMATION ACT PROGRAM § 293.2 Policy. It is NIMA policy that: (a) Agency records that, if disclosed, would cause no foreseeable harm to an interest...

  16. 5 CFR 293.307 - Disposition of folders of former Federal employees.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Disposition of folders of former Federal employees. 293.307 Section 293.307 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.307 Disposition of folders of former Federal...

  17. 5 CFR 293.304 - Maintenance and content of folder.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Maintenance and content of folder. 293.304 Section 293.304 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.304 Maintenance and content of folder. The head of each...

  18. 5 CFR 293.304 - Maintenance and content of folder.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Maintenance and content of folder. 293.304 Section 293.304 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.304 Maintenance and content of folder. The head of each...

  19. 5 CFR 293.105 - Restrictions on collection and use of information.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Restrictions on collection and use of information. 293.105 Section 293.105 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.105 Restrictions on...

  20. 8 CFR 293.3 - Simple interest table.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 8 Aliens and Nationality 1 2010-01-01 2010-01-01 false Simple interest table. 293.3 Section 293.3 Aliens and Nationality DEPARTMENT OF HOMELAND SECURITY IMMIGRATION REGULATIONS DEPOSIT OF AND INTEREST ON CASH RECEIVED TO SECURE IMMIGRATION BONDS § 293.3 Simple interest table. Following is a simple interest...

  1. 36 CFR 293.8 - Permanent structures and commercial services.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 36 Parks, Forests, and Public Property 2 2014-07-01 2014-07-01 false Permanent structures and commercial services. 293.8 Section 293.8 Parks, Forests, and Public Property FOREST SERVICE, DEPARTMENT OF AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.8 Permanent structures and commercial services. Motels, summer...

  2. 36 CFR 293.8 - Permanent structures and commercial services.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 36 Parks, Forests, and Public Property 2 2013-07-01 2013-07-01 false Permanent structures and commercial services. 293.8 Section 293.8 Parks, Forests, and Public Property FOREST SERVICE, DEPARTMENT OF AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.8 Permanent structures and commercial services. Motels, summer...

  3. 5 CFR 293.507 - Maintenance and content of the Employee Medical Folder.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Maintenance and content of the Employee Medical Folder. 293.507 Section 293.507 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.507 Maintenance and...

  4. 5 CFR 293.504 - Composition of, and access to, the Employee Medical File System.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Employee Medical File System. 293.504 Section 293.504 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.504 Composition of, and access to, the Employee Medical File System. (a) All employee occupational medical records...

  5. 5 CFR 293.507 - Maintenance and content of the Employee Medical Folder.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Maintenance and content of the Employee Medical Folder. 293.507 Section 293.507 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.507 Maintenance and...

  6. 5 CFR 293.504 - Composition of, and access to, the Employee Medical File System.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Employee Medical File System. 293.504 Section 293.504 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.504 Composition of, and access to, the Employee Medical File System. (a) All employee occupational medical records...

  7. 5 CFR 293.506 - Ownership of the Employee Medical Folder.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Ownership of the Employee Medical Folder. 293.506 Section 293.506 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.506 Ownership of the Employee Medical...

  8. 5 CFR 293.506 - Ownership of the Employee Medical Folder.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Ownership of the Employee Medical Folder. 293.506 Section 293.506 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.506 Ownership of the Employee Medical...

  9. 5 CFR 293.506 - Ownership of the Employee Medical Folder.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Ownership of the Employee Medical Folder. 293.506 Section 293.506 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.506 Ownership of the Employee Medical...

  10. 5 CFR 293.507 - Maintenance and content of the Employee Medical Folder.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Maintenance and content of the Employee Medical Folder. 293.507 Section 293.507 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.507 Maintenance and...

  11. 5 CFR 293.504 - Composition of, and access to, the Employee Medical File System.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Employee Medical File System. 293.504 Section 293.504 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.504 Composition of, and access to, the Employee Medical File System. (a) All employee occupational medical records...

  12. 5 CFR 293.504 - Composition of, and access to, the Employee Medical File System.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Employee Medical File System. 293.504 Section 293.504 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.504 Composition of, and access to, the Employee Medical File System. (a) All employee occupational medical records...

  13. 5 CFR 293.507 - Maintenance and content of the Employee Medical Folder.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Maintenance and content of the Employee Medical Folder. 293.507 Section 293.507 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.507 Maintenance and...

  14. 5 CFR 293.507 - Maintenance and content of the Employee Medical Folder.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Maintenance and content of the Employee Medical Folder. 293.507 Section 293.507 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.507 Maintenance and...

  15. 5 CFR 293.506 - Ownership of the Employee Medical Folder.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Ownership of the Employee Medical Folder. 293.506 Section 293.506 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.506 Ownership of the Employee Medical...

  16. 5 CFR 293.504 - Composition of, and access to, the Employee Medical File System.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Employee Medical File System. 293.504 Section 293.504 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.504 Composition of, and access to, the Employee Medical File System. (a) All employee occupational medical records...

  17. 5 CFR 293.506 - Ownership of the Employee Medical Folder.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Ownership of the Employee Medical Folder. 293.506 Section 293.506 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.506 Ownership of the Employee Medical...

  18. 43 CFR 29.3 - Fund administration.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 43 Public Lands: Interior 1 2011-10-01 2011-10-01 false Fund administration. 29.3 Section 29.3 Public Lands: Interior Office of the Secretary of the Interior TRANS-ALASKA PIPELINE LIABILITY FUND § 29.3 Fund administration. (a) The Fund shall be administered by a Board of Trustees designated by the...

  19. Nano-twin Mediated Plasticity in Carbon-containing FeNiCoCrMn High Entropy Alloys

    DOE PAGES

    Wu, Zhenggang; Bei, Hongbin; Parish, Chad M

    2015-06-14

    Equiatomic FeNiCoCrMn alloy has been reported to exhibit promising strength and ductility at cryogenic temperature and deformation mediated by nano-twining appeared to be one of the main reasons. We use the FeNiCoCrMn alloy as a base alloy to seek further improvement of its mechanical properties by alloying additional elements, i.e., interstitial carbon. Moreover, the effects of carbon on microstructures, mechanical properties and twinning activities were investigated in two different temperatures (77 and 293 K). With addition of 0.5 at% C, the high entropy alloy still remains entirely single phase face-centered cubic (FCC) crystal structure. We found that these materials canmore » be cold rolled and recrystallized to produce a microstructure with equiaxed grains. Both strain hardening rate and strength are enhanced while high uniform elongations to fracture (~70% at 77 K and ~40% at 293 K) are still maintained. The increased strain hardening and strength could be caused by the promptness of deformation twinning in C-containing high entropy alloys.« less

  20. Mapping analysis of scaffold/matrix attachment regions (s/MARs) from two different mammalian cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pilus, Nur Shazwani Mohd; Ahmad, Azrin; Yusof, Nurul Yuziana Mohd

    Scaffold/matrix attachment regions (S/MARs) are potential element that can be integrated into expression vector to increase expression of recombinant protein. Many studies on S/MAR have been done but none has revealed the distribution of S/MAR in a genome. In this study, we have isolated S/MAR sequences from HEK293 and Chinese hamster ovary cell lines (CHO DG44) using two different methods utilizing 2 M NaCl and lithium-3,5-diiodosalicylate (LIS). The isolated S/MARs were sequenced using Next Generation Sequencing (NGS) platform. Based on reference mapping analysis against human genome database, a total of 8,994,856 and 8,412,672 contigs of S/MAR sequences were retrieved frommore » 2M NaCl and LIS extraction of HEK293 respectively. On the other hand, reference mapping analysis of S/MAR derived from CHO DG44 against our own CHO DG44 database have generated a total of 7,204,348 and 4,672,913 contigs from 2 M NaCl and LIS extraction method respectively.« less

  1. Production and purification of lentiviral vectors generated in 293T suspension cells with baculoviral vectors.

    PubMed

    Lesch, H P; Laitinen, A; Peixoto, C; Vicente, T; Makkonen, K-E; Laitinen, L; Pikkarainen, J T; Samaranayake, H; Alves, P M; Carrondo, M J T; Ylä-Herttuala, S; Airenne, K J

    2011-06-01

    Lentivirus can be engineered to be a highly potent vector for gene therapy applications. However, generation of clinical grade vectors in enough quantities for therapeutic use is still troublesome and limits the preclinical and clinical experiments. As a first step to solve this unmet need we recently introduced a baculovirus-based production system for lentiviral vector (LV) production using adherent cells. Herein, we have adapted and optimized the production of these vectors to a suspension cell culture system using recombinant baculoviruses delivering all elements required for a safe latest generation LV preparation. High-titer LV stocks were achieved in 293T cells grown in suspension. Produced viruses were accurately characterized and the functionality was also tested in vivo. Produced viruses were compared with viruses produced by calcium phosphate transfection method in adherent cells and polyethylenimine transfection method in suspension cells. Furthermore, a scalable and cost-effective capture purification step was developed based on a diethylaminoethyl monolithic column capable of removing most of the baculoviruses from the LV pool with 65% recovery.

  2. Chemical Dissolution of Simulant FCA Cladding and Plates

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Daniel, G.; Pierce, R.; O'Rourke, P.

    The Savannah River Site (SRS) has received some fast critical assembly (FCA) fuel from the Japan Atomic Energy Agency (JAEA) for disposition. Among the JAEA FCA fuel are approximately 7090 rectangular Stainless Steel clad fuel elements. Each element has an internal Pu-10.6Al alloy metal wafer. The thickness of each element is either 1/16 inch or 1/32 inch. The dimensions of each element ranges from 2 inches x 1 inch to 2 inches x 4 inches. This report discusses the potential chemical dissolution of the FCA clad material or stainless steel. This technology uses nitric acid-potassium fluoride (HNO 3-KF) flowsheets ofmore » H-Canyon to dissolve the FCA elements from a rack of materials. Historically, dissolution flowsheets have aimed to maximize Pu dissolution rates while minimizing stainless steel dissolution (corrosion) rates. Because the FCA cladding is made of stainless steel, this work sought to accelerate stainless steel dissolution.« less

  3. Design of pneumatic proportional flow valve type 5/3

    NASA Astrophysics Data System (ADS)

    Laski, P. A.; Pietrala, D. S.; Zwierzchowski, J.; Czarnogorski, K.

    2017-08-01

    In this paper the 5/3-way pneumatic, proportional flow valve was designed and made. Stepper linear actuator was used to move the spool. The valve is controlled by the controlled based on a AVR microcontroller. Virtual model of the valve was created in CAD. The real element was made based on a standard 5/3-way manually actuated valve with hand lever, which was dismounted and replaced by linear stepper motor. All the elements was mounted in a specially made housing. The controller consists of microcontroller Atmega16, integrated circuit L293D, display, two potentiometers, three LEDs and six buttons. Series of research was also conducted. Simulation research were performed using CFD by the Flow Simulation addition to SolidWorks. During the experiments the valve characteristics of flow and pressure was determined.

  4. 5 CFR 293.403 - Contents of employee performance files.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Contents of employee performance files. 293.403 Section 293.403 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.403 Contents of employee performance files. (a) A decision on what constitutes ...

  5. 28 CFR 29.3 - Administration by the Bureau of Justice Assistance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Administration by the Bureau of Justice Assistance. 29.3 Section 29.3 Judicial Administration DEPARTMENT OF JUSTICE MOTOR VEHICLE THEFT PREVENTION ACT REGULATIONS § 29.3 Administration by the Bureau of Justice Assistance. The Director of the Bureau...

  6. 8 CFR 293.2 - Interest rate.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 8 Aliens and Nationality 1 2010-01-01 2010-01-01 false Interest rate. 293.2 Section 293.2 Aliens... CASH RECEIVED TO SECURE IMMIGRATION BONDS § 293.2 Interest rate. The Secretary of the Treasury has determined that effective from date of deposit occurring after April 27, 1966, the interest rate shall be 3...

  7. 5 CFR 293.305 - Type of folder to be used.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Type of folder to be used. 293.305 Section 293.305 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.305 Type of folder to be used. Each agency shall use only...

  8. 8 CFR 293.2 - Interest rate.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 8 Aliens and Nationality 1 2014-01-01 2014-01-01 false Interest rate. 293.2 Section 293.2 Aliens... CASH RECEIVED TO SECURE IMMIGRATION BONDS § 293.2 Interest rate. The Secretary of the Treasury has determined that effective from date of deposit occurring after April 27, 1966, the interest rate shall be 3...

  9. 8 CFR 293.2 - Interest rate.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 8 Aliens and Nationality 1 2012-01-01 2012-01-01 false Interest rate. 293.2 Section 293.2 Aliens... CASH RECEIVED TO SECURE IMMIGRATION BONDS § 293.2 Interest rate. The Secretary of the Treasury has determined that effective from date of deposit occurring after April 27, 1966, the interest rate shall be 3...

  10. 8 CFR 293.2 - Interest rate.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 8 Aliens and Nationality 1 2011-01-01 2011-01-01 false Interest rate. 293.2 Section 293.2 Aliens... CASH RECEIVED TO SECURE IMMIGRATION BONDS § 293.2 Interest rate. The Secretary of the Treasury has determined that effective from date of deposit occurring after April 27, 1966, the interest rate shall be 3...

  11. 8 CFR 293.2 - Interest rate.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 8 Aliens and Nationality 1 2013-01-01 2013-01-01 false Interest rate. 293.2 Section 293.2 Aliens... CASH RECEIVED TO SECURE IMMIGRATION BONDS § 293.2 Interest rate. The Secretary of the Treasury has determined that effective from date of deposit occurring after April 27, 1966, the interest rate shall be 3...

  12. 5 CFR 293.509 - Use of existing Employee Medical Folders upon transfer or reemployment.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Use of existing Employee Medical Folders upon transfer or reemployment. 293.509 Section 293.509 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.509 Use of...

  13. 5 CFR 293.508 - Type of folder to be used.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Type of folder to be used. 293.508 Section 293.508 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.508 Type of folder to be used. Each agency must...

  14. 5 CFR 293.508 - Type of folder to be used.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Type of folder to be used. 293.508 Section 293.508 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.508 Type of folder to be used. Each agency must...

  15. 5 CFR 293.509 - Use of existing Employee Medical Folders upon transfer or reemployment.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Use of existing Employee Medical Folders upon transfer or reemployment. 293.509 Section 293.509 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.509 Use of...

  16. 5 CFR 293.508 - Type of folder to be used.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Type of folder to be used. 293.508 Section 293.508 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.508 Type of folder to be used. Each agency must...

  17. 5 CFR 293.508 - Type of folder to be used.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Type of folder to be used. 293.508 Section 293.508 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.508 Type of folder to be used. Each agency must...

  18. 5 CFR 293.509 - Use of existing Employee Medical Folders upon transfer or reemployment.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Use of existing Employee Medical Folders upon transfer or reemployment. 293.509 Section 293.509 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.509 Use of...

  19. 5 CFR 293.509 - Use of existing Employee Medical Folders upon transfer or reemployment.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Use of existing Employee Medical Folders upon transfer or reemployment. 293.509 Section 293.509 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.509 Use of...

  20. 5 CFR 293.509 - Use of existing Employee Medical Folders upon transfer or reemployment.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Use of existing Employee Medical Folders upon transfer or reemployment. 293.509 Section 293.509 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.509 Use of...

  1. 5 CFR 293.508 - Type of folder to be used.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Type of folder to be used. 293.508 Section 293.508 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Medical File System Records § 293.508 Type of folder to be used. Each agency must...

  2. Cooperation of NAD(P)H:quinone oxidoreductase 1 and UDP-glucuronosyltransferases reduces menadione cytotoxicity in HEK293 cells.

    PubMed

    Nishiyama, Takahito; Izawa, Tadashi; Usami, Mami; Ohnuma, Tomokazu; Ogura, Kenichiro; Hiratsuka, Akira

    2010-04-09

    Previous studies have shown that NAD(P)H:quinone oxidoreductase 1 (NQO1) plays an important role in the detoxification of menadione (2-methyl-1,4-naphthoquinone, also known as vitamin K3). However, menadiol (2-methyl-1,4-naphthalenediol) formed from menadione by NQO1-mediated reduction continues to be an unstable substance, which undergoes the reformation of menadione with concomitant formation of reactive oxygen species (ROS). Hence, we focused on the roles of phase II enzymes, with particular attention to UDP-glucuronosyltransferases (UGTs), in the detoxification process of menadione. In this study, we established an HEK293 cell line stably expressing NQO1 (HEK293/NQO1) and HEK293/NQO1 cell lines with doxycycline (DOX)-regulated expression of UGT1A6 (HEK293/NQO1/UGT1A6) and UGT1A10 (HEK293/NQO1/UGT1A10), and evaluated the role of NQO1 and UGTs against menadione-induced cytotoxicity. Our results differed from those of previous studies. HEK293/NQO1 was the most sensitive cell line to menadione cytotoxicity among cell lines established in this study. These phenomena were also observed in HEK293/NQO1/UGT1A6 and HEK293/NQO1/UGT1A10 cells in which the expression of UGT was suppressed by DOX treatment. On the contrary, HEK293/NQO1/UGT1A6 and HEK293/NQO1/UGT1A10 cells without DOX treatment were resistant to menadione-induced cytotoxicity. These results demonstrated that NQO1 is not a detoxification enzyme for menadione and that UGT-mediated glucuronidation of menadiol is the most important detoxification process. Copyright 2009 Elsevier Inc. All rights reserved.

  3. Limitations to the development of recombinant human embryonic kidney 293E cells using glutamine synthetase-mediated gene amplification: Methionine sulfoximine resistance.

    PubMed

    Yu, Da Young; Noh, Soo Min; Lee, Gyun Min

    2016-08-10

    To investigate the feasibility of glutamine synthetase (GS)-mediated gene amplification in HEK293 cells for the high-level stable production of therapeutic proteins, HEK293E cells were transfected by the GS expression vector containing antibody genes and were selected at various methionine sulfoximine (MSX) concentrations in 96-well plates. For a comparison, CHOK1 cells were transfected by the same GS expression vector and selected at various MSX concentrations. Unlike CHOK1 cells, HEK293E cells producing high levels of antibodies were not selected at all. For HEK293E cells, the number of wells with the cell pool did not decrease with an increase in the concentration of MSX up to 500μM MSX. A q-RT-PCR analysis confirmed that the antibody genes in the HEK293E cells, unlike the CHOK1 cells, were not amplified after increasing the MSX concentration. It was found that the GS activity in HEK293E cells was much higher than that in CHOK1 cells (P<0.05). In a glutamine-free medium, the GS activity of HEK293E cells was approximately 4.8 times higher than that in CHOK1 cells. Accordingly, it is inferred that high GS activity of HEK293E cells results in elevated resistance to MSX and therefore hampers GS-mediated gene amplification by MSX. Thus, in order to apply the GS-mediated gene amplification system to HEK293 cells, the endogenous GS expression level in HEK293 cells needs to be minimized by knock-out or down-regulation methods. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Purification of Recombinant Ebola Virus Glycoprotein and VP40 from a Human Cell Line

    DTIC Science & Technology

    2017-01-01

    from a human cell line. Plasmids coding for the expression of these proteins were transiently transfected into human embryonic kidney cells 293 and...protein expression. Expi293F cells were derived from the line of human embryonic kidney cells 293 (i.e., HEK293 cells), and they were grown in a

  5. 33 CFR 334.293 - Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. 334.293 Section 334.293 Navigation and... RESTRICTED AREA REGULATIONS § 334.293 Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. (a) The area. (1) The waters within an area beginning at a point on the...

  6. 33 CFR 334.293 - Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. 334.293 Section 334.293 Navigation and... RESTRICTED AREA REGULATIONS § 334.293 Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. (a) The area. (1) The waters within an area beginning at a point on the...

  7. 33 CFR 334.293 - Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. 334.293 Section 334.293 Navigation and... RESTRICTED AREA REGULATIONS § 334.293 Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. (a) The area. (1) The waters within an area beginning at a point on the...

  8. 33 CFR 334.293 - Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. 334.293 Section 334.293 Navigation and... RESTRICTED AREA REGULATIONS § 334.293 Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. (a) The area. (1) The waters within an area beginning at a point on the...

  9. 33 CFR 334.293 - Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. 334.293 Section 334.293 Navigation and... RESTRICTED AREA REGULATIONS § 334.293 Elizabeth River, Craney Island Refueling Pier Restricted Area, Portsmouth VA; naval restricted area. (a) The area. (1) The waters within an area beginning at a point on the...

  10. Computer Resources Handbook for Flight Critical Systems.

    DTIC Science & Technology

    1985-01-01

    8183 UNCLASSIFIED F /G 1 /3 NLllEEEEEEEEEEEE Illll lflflflflfl Illlfll..lflll EIEEIhEEIIIIII llElfllfllflfllfll .flflflflflflflflflflflll llEEEEEEllEII...Requiring Simulation .... ............. ... 114 6-4 Typical Transition Elements of Care III. ...... ............ 116 D- 1 F -16 Digital Flight Control...following forms: 1 . By reference to MIL- F -9490(for flight control systems); 2. By reference, with tailoring if required, to a Federal Aviation Regulation

  11. Crystallization and preliminary X-ray diffraction analyses of the redox-controlled complex of terminal oxygenase and ferredoxin components in the Rieske nonhaem iron oxygenase carbazole 1,9a-dioxygenase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Matsuzawa, Jun; Aikawa, Hiroki; Umeda, Takashi

    2014-09-25

    A crystal was obtained of the complex between reduced terminal oxygenase and oxidized ferredoxin components of carbazole 1,9a-dioxygenase. The crystal belonged to space group P2{sub 1} and diffracted to 2.25 Å resolution. The initial reaction in bacterial carbazole degradation is catalyzed by carbazole 1,9a-dioxygenase, which consists of terminal oxygenase (Oxy), ferredoxin (Fd) and ferredoxin reductase components. The electron-transfer complex between reduced Oxy and oxidized Fd was crystallized at 293 K using the hanging-drop vapour-diffusion method with PEG 3350 as the precipitant under anaerobic conditions. The crystal diffracted to a maximum resolution of 2.25 Å and belonged to space group P2{submore » 1}, with unit-cell parameters a = 97.3, b = 81.6, c = 116.2 Å, α = γ = 90, β = 100.1°. The V{sub M} value is 2.85 Å{sup 3} Da{sup −1}, indicating a solvent content of 56.8%.« less

  12. 25 CFR 293.15 - When does an approved or considered-to-have-been-approved compact or amendment take effect?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... compact or amendment take effect? 293.15 Section 293.15 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR ECONOMIC ENTERPRISES CLASS III TRIBAL STATE GAMING COMPACT PROCESS § 293.15 When does an approved or considered-to-have-been-approved compact or amendment take effect? (a) An approved or...

  13. Construction of recombinant FGFR1 containing full-length gene and its potential application.

    PubMed

    Zhou, Yali; Luo, Wenjuan; Zheng, Lei; Li, Miao; Zhang, Yanmin

    2010-07-01

    FGFR1, one of the four fibroblast growth factor receptors, has been found to be over-expressed in many cancers. In this study, a full-length expression plasmid for FGFR1 was obtained by fragment amplification. The amplified PCR product was then digested and inserted into the pcDNA3.1(+) vector. A recombinant eukaryotic expression vector containing the complete CDS region of FGFR1 was successfully constructed. After it was transfected to Hek293 cell, the expression of the FGFR1 receptor in recombinant Hek293/FGFR1 was 18 times higher than that of Hek293 cell. The biological activities of high expression FGFR1 cell (Hek293/FGFR1) were verified by FCM, immunofluorescent, RT-PCR, western blot and cell cycle analysis. Then, Hek293/FGFR1 was used to screen taspine with cell membrane chromatography (CMC). Finally, we analyzed the effects of taspine on Hek293/FGFR1 cell and MCF-7 cell. In conclusion, Hek293/FGFR1 was successfully constructed. The results demonstrate that taspine can down-regulate phosphorylation of FGFR1 and ERK, and inhibit Hek293/FGFR1 and MCF-7 cell proliferation. Copyright 2010 Elsevier Inc. All rights reserved.

  14. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kang, Kyungsu; Lee, Kyung-Mi; Yoo, Ji-Hye

    Graphical abstract: Schematic diagram of the possible molecular mechanism underlying the inhibition of the Wnt/{beta}-catenin signaling pathway and the induction of G0/G1-phase arrest by gomisins J and N, derived from the fruits of S. chinensis, in HCT116 human colon cancer cells. Highlights: Black-Right-Pointing-Pointer Gomisins J and N inhibited Wnt/{beta}-catenin signaling pathway in HCT116 cells. Black-Right-Pointing-Pointer Gomisins J and N disrupted the binding of {beta}-catenin to specific DNA sequences, TBE. Black-Right-Pointing-Pointer Gomisins J and N inhibited the HCT116 cell proliferation through G0/G1 phase arrest. Black-Right-Pointing-Pointer Gomisins J and N inhibited the expression of Cyc D1, a Wnt/{beta}-catenin target gene. -- Abstract:more » Here, we report that gomisin J and gomisin N, dibenzocyclooctadiene type lignans isolated from Schisandra chinensis, inhibit Wnt/{beta}-catenin signaling in HCT116 cells. Gomisins J and N appear to inhibit Wnt/{beta}-catenin signaling by disrupting the interaction between {beta}-catenin and its specific target DNA sequences (TCF binding elements, TBE) rather than by altering the expression of the {beta}-catenin protein. Gomisins J and N inhibit HCT116 cell proliferation by arresting the cell cycle at the G0/G1 phase. The G0/G1 phase arrest induced by gomisins J and N appears to be caused by a decrease in the expression of Cyclin D1, a representative target gene of the Wnt/{beta}-catenin signaling pathway, as well as Cdk2, Cdk4, and E2F-1. Therefore, gomisins J and N, the novel Wnt/{beta}-catenin inhibitors discovered in this study, may serve as potential agents for the prevention and treatment of human colorectal cancers.« less

  15. Activation of RNA polymerase III transcription of human Alu repetitive elements by adenovirus type 5: Requirement for the E1b 58-Kilodalton protein and the products of E4 open reading frames 3 and 6

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Panning, B.; Smiley, J.R.

    1993-06-01

    Alu elements are the single most abundant class of dispersed repeated sequences in the human genome, comprising 5-10% of the mass of human DNA. This report demonstrates that Ad5 infection strongly stimulates Pol III transcription of human Alu elements in HeLa and 293 cells. In contrast to the cases of Ad5-induced Pol III transcriptional activation, this process requires the E1b 58-kDa protein and the products of E4 open reading frames (ORFs) 3 and 6 in addition to the E1a 289-residue product. These findings suggest novel regulatory properties of the Ad5 E1b and E4 proteins and raise the possibility that analogousmore » cellular trans-acting factors serve to modulate Alu expression in vivo.« less

  16. Chikusetsusaponin IVa methyl ester induces cell cycle arrest by the inhibition of nuclear translocation of β-catenin in HCT116 cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lee, Kyung-Mi; Yun, Ji Ho; Lee, Dong Hwa

    2015-04-17

    We demonstrate that chikusetsusaponin IVa methyl ester (CME), a triterpenoid saponin from the root of Achyranthes japonica, has an anticancer activity. We investigate its molecular mechanism in depth in HCT116 cells. CME reduces the amount of β-catenin in nucleus and inhibits the binding of β-catenin to specific DNA sequences (TCF binding elements, TBE) in target gene promoters. Thus, CME appears to decrease the expression of cell cycle regulatory proteins such as Cyclin D1, as a representative target for β-catenin, as well as CDK2 and CDK4. As a result of the decrease of the cell cycle regulatory proteins, CME inhibits cellmore » proliferation by arresting the cell cycle at the G0/G1 phase. Therefore, we suggest that CME as a novel Wnt/β-catenin inhibitor can be a putative agent for the treatment of colorectal cancers. - Highlights: • CME inhibits cell proliferation in HCT116 cells. • CME increases cell cycle arrest at G0/G1 phase and apoptosis. • CME attenuates cyclin D1 and regulates cell cycle regulatory proteins. • CME inhibits β-catenin translocation to nucleus.« less

  17. [Determination of heavy metals in four traditional Chinese medicines by ICP-MS].

    PubMed

    Wen, Hui-Min; Chen, Xiao-Hui; Dong, Ting-Xia; Zhan, Hua-Qiang; Bi, Kai-Shun

    2006-08-01

    To establish a ICP-MS method for the determination of heavy metals, including As, Hg, Pb, Cd, in four traditional Chinese medicines. The samples were digested by closed-versel microwave. The four heavy metals were directly analyzed by ICP-MS. Select internal standard element in for the method by which the analyse signal drife is corrected by the signal of another element (internal standard elements) added to both the standard solution and sample. For all of the analyzed heary methals, the correlative coefficient of the calibration curves was over 0.999 2. The recovery rates of the procedure were 97.5%-108.0%, and its RSD was lower than 11.6%. This method was convenient, quick-acquired, accurate and highly sensitive. The method can be used for the quality control of trace elements in traditional Chinese medicines and for the contents determination of traditional Chinese medicines from different habitats and species.

  18. Applying the silver-tube introduction method for thermal conversion elemental analyses and a new δ2H value for NBS 22 oil

    USGS Publications Warehouse

    Coplen, Tyler B.; Qi, Haiping

    2010-01-01

    The δ2HVSMOW–SLAP value of total hydrogen of the international measurement standard NBS 22 oil was determined by a new method of sealing water in silver tubes for use in a thermal conversion elemental analysis (TC/EA) reduction unit. The isotopic fractionation of water due to evaporation is virtually non-existent in this silver-tube method. A new value for the δ2HVSMOW–SLAP of NBS 22 oil, calibrated with isotopic reference waters, was determined to be −116.9 ± 0.8‰ (1σ and n = 31).

  19. Ba 2TeO: A new layered oxytelluride

    DOE PAGES

    Besara, T.; Ramirez, D.; Sun, J.; ...

    2015-02-01

    For single crystals of the new semiconducting oxytelluride phase, Ba 2TeO, we synthesized from barium oxide powder and elemental tellurium in a molten barium metal flux. Ba 2TeO crystallizes in tetragonal symmetry with space group P4/nmm (#129), a=5.0337(1) Å, c=9.9437(4) Å, Z=2. The crystals were characterized by single crystal x-ray diffraction, heat capacity and optical measurements. Moreover, the optical measurements along with electronic band structure calculations indicate semiconductor behavior with a band gap of 2.93 eV. Resistivity measurements show that Ba 2TeO is highly insulating.

  20. Determination of trace elements in honey from different regions in Rio de Janeiro State (Brazil) by total reflection X-ray fluorescence.

    PubMed

    Ribeiro, Roberta de Oliveira Resende; Mársico, Eliane Teixeira; de Jesus, Edgar Francisco Oliveira; da Silva Carneiro, Carla; Júnior, Carlos Adam Conte; de Almeida, Eduardo; Filho, Virgílio Franco do Nascimento

    2014-04-01

    Trace and minor elements in Brazilian honey were analyzed by total reflection X-ray fluorescence spectroscopy. Up to 12 elements (K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br, and Sr) were detected in 160 samples of honey from 4 regions of Rio de Janeiro State (Barra Mansa, Teresópolis, northern and southern Nova Friburgo). The results showed the samples from Teresópolis had higher rates of essential and nonessential elements than samples from the other regions, except for Ni. K and Ca were the most abundant elements in all samples, in the range of 116.5 to 987.0 μg g(-1) . Ni, Cu, Zn, Se, and Sr were identified in small concentrations (0.01 to 12.08 μg g(-1) ) in all samples, indicating a low level of contamination in all the regions. © 2014 Institute of Food Technologists®

  1. 5 CFR 293.303 - Ownership of folder.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Ownership of folder. 293.303 Section 293.303 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL... the records of, the Office of Personnel Management (the Office). ...

  2. 5 CFR 293.303 - Ownership of folder.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Ownership of folder. 293.303 Section 293.303 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL... the records of, the Office of Personnel Management (the Office). ...

  3. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Togi, Sumihito; Nakasuji, Misa; Muromoto, Ryuta

    Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded latency-associated nuclear antigen (LANA), which interacts with cellular proteins, plays a central role in modification of viral and/or cellular gene expression. Here, we show that LANA associates with glucocorticoid receptor (GR), and that LANA enhances the transcriptional activity of GR. Co-immunoprecipitation revealed a physical interaction between LANA and GR in transiently transfected 293T and HeLa cells. In human B-lymphoma cells, LANA overexpression enhanced GR activity and cell growth suppression following glucocorticoid stimulation. Furthermore, confocal microscopy showed that activated GR was bound to LANA and accumulated in the nucleus, leading to an increase in binding of activatedmore » GR to the glucocorticoid response element of target genes. Taken together, KSHV-derived LANA acts as a transcriptional co-activator of GR. Our results might suggest a careful use of glucocorticoids in the treatment of patients with KSHV-related malignancies such as Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman disease. - Highlights: • KSHV-LANA enhances the transcriptional activity of GR in 293T and HeLa cells. • KSHV-LANA physically associates with GR. • KSHV-LANA enhances GR activation and cell growth suppression in human B-lymphocytes. • KSHV-LANA influences the nuclear retention and DNA binding activity of GR.« less

  4. 5 CFR 293.103 - Recordkeeping standards.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 293.103 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.103 Recordkeeping standards. (a) The head..., processing, use, or maintenance of personnel records are informed of pertinent recordkeeping regulations and...

  5. An analysis of state legislation on community trails.

    PubMed

    Eyler, Amy; Lankford, Tina; Chriqui, Jamie; Evenson, Kelly R; Kruger, Judy; Tompkins, Nancy; Voorhees, Carolyn; Zieff, Susan; Aytur, Semra; Brownson, Ross

    2010-03-01

    Trails provide opportunities for recreation, transportation and activity. The purpose of this article is to describe state legislation related to community trails, to analyze legislation content, and to evaluate legislation on inclusion of evidence-informed elements. State trail legislation from 2001 to 2008 was identified using online legislative databases. An analysis of evidence-informed elements included in the legislation was conducted. These elements included: funding, liability, accessibility, connectivity, and maintenance. Of the total 991 trail bills, 516 (52.0%) were appropriations bills, of which 167 (32.2%) were enacted. We analyzed 475 (48%) nonappropriation trail bills of which 139 (29.3%) were enacted. The percentage of enactment of appropriations bills decreased over time while enactment of nonappropriations trail bills increased. Over half of the nonappropriations trail bills included at least 1 evidence-informed element, most commonly funding. Few bills contained liability, connectivity, accessibility, or maintenance. There is opportunity for providing evidence-informed information to policy-makers to potentially influence bill content. The number of bills with a funding element demonstrates that fiscal support for trails is an important policy lever that state legislatures may use to support trails. Lastly, trails should be considered in over-all state-level physical activity legislation to provide opportunities for communities to be active.

  6. Smad4-Mediated Signaling Inhibits Intestinal Neoplasia by Inhibiting Expression of β-Catenin

    PubMed Central

    Freeman, Tanner J.; Smith, J. Joshua; Chen, Xi; Washington, M. Kay; Roland, Joseph T.; Means, Anna L.; Eschrich, Steven A.; Yeatman, Timothy J.; Deane, Natasha G.; Beauchamp, R. Daniel

    2012-01-01

    Background & Aims Mutational inactivation of APC is an early event in colorectal cancer (CRC) progression that affects the stability and increases the activity of β-catenin, a mediator of Wnt signaling. CRC progression also involves inactivation of signaling via transforming growth factor (TGF)β and bone morphenogenic protein (BMP), which are tumor suppressors. However, the interactions between these pathways are not clear. We investigated the effects of loss of the transcription factor Smad4 loss on levels of β-catenin mRNA and Wnt signaling. Methods We used microarray analysis to associate levels of Smad4 and β-catenin mRNA in colorectal tumor samples from 250 patients. We performed oligonucleotide-mediated knockdown of Smad4 in human embryonic kidney (HEK293T) and in HCT116 colon cancer cells and transgenically expressed Smad4 in SW480 colon cancer cells. We analyzed adenomas from (APCΔ1638/+) and (APCΔ1638/+)x(K19CreERT2Smad4lox/lox) mice using laser-capture microdissection. Results In human CRC samples, reduced levels of Smad4 correlated with increased levels of β-catenin mRNA. In Smad4-depleted cell lines, levels of β-catenin mRNA and Wnt signaling increased. Inhibition of BMP or depletion of Smad4 in HEK293T cells increased binding of RNA polymerase II to the β-catenin gene. Expression of Smad4 in SW480 cells reduced Wnt signaling and levels of β-catenin mRNA. In mice with heterozygous disruption of Apc(APCΔ1638/+), Smad4-deficient intestinal adenomas had increased levels of β-catenin mRNA and expression of Wnt target genes, compared with adenomas from APCΔ1638/+mice that expressed Smad4. Conclusions Transcription of β-catenin is inhibited by BMP signaling to Smad4. These findings provide important information about the interaction among TGF-β, BMP, and Wnt signaling pathways in CRC progression. PMID:22115830

  7. 5 CFR 293.103 - Recordkeeping standards.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... requirements of the Office of Personnel Management and the agency. Authority to maintain personnel records does... 293.103 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.103 Recordkeeping standards. (a) The head...

  8. 5 CFR 293.103 - Recordkeeping standards.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... requirements of the Office of Personnel Management and the agency. Authority to maintain personnel records does... 293.103 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.103 Recordkeeping standards. (a) The head...

  9. 5 CFR 293.103 - Recordkeeping standards.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... requirements of the Office of Personnel Management and the agency. Authority to maintain personnel records does... 293.103 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.103 Recordkeeping standards. (a) The head...

  10. 5 CFR 293.103 - Recordkeeping standards.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... requirements of the Office of Personnel Management and the agency. Authority to maintain personnel records does... 293.103 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.103 Recordkeeping standards. (a) The head...

  11. Solubility of carbon monoxide in n-hexane between 293 and 473 K and CO pressures up to 200 bar

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Koelliker, R.; Thies, H.

    The solubility of carbon monoxide, CO, in n-hexane was measured at 293, 323, 373, 423, and 473 K for CO partial pressures up to 200 bar. The enthalpy of solution was calculated between 293 and 473 K. Using the Krichevsky-Ilinskaya equation of state, the solubility of CO in n-hexane can be calculated between 293 and 423 K for CO partial pressures up to 200 bar with an accuracy better than 5%.

  12. Analysis of cis and trans Requirements for DNA Replication at the Right-End Hairpin of the Human Bocavirus 1 Genome

    PubMed Central

    Shen, Weiran; Deng, Xuefeng; Zou, Wei; Engelhardt, John F.; Yan, Ziying

    2016-01-01

    ABSTRACT Parvoviruses are single-stranded DNA viruses that use the palindromic structures at the ends of the viral genome for their replication. The mechanism of parvovirus replication has been studied mostly in the dependoparvovirus adeno-associated virus 2 (AAV2) and the protoparvovirus minute virus of mice (MVM). Here, we used human bocavirus 1 (HBoV1) to understand the replication mechanism of bocaparvovirus. HBoV1 is pathogenic to humans, causing acute respiratory tract infections, especially in young children under 2 years old. By using the duplex replicative form of the HBoV1 genome in human embryonic kidney 293 (HEK293) cells, we identified the HBoV1 minimal replication origin at the right-end hairpin (OriR). Mutagenesis analyses confirmed the putative NS1 binding and nicking sites within the OriR. Of note, unlike the large nonstructural protein (Rep78/68 or NS1) of other parvoviruses, HBoV1 NS1 did not specifically bind OriR in vitro, indicating that other viral and cellular components or the oligomerization of NS1 is required for NS1 binding to the OriR. In vivo studies demonstrated that residues responsible for NS1 binding and nicking are within the origin-binding domain. Further analysis identified that the small nonstructural protein NP1 is required for HBoV1 DNA replication at OriR. NP1 and other viral nonstructural proteins (NS1 to NS4) colocalized within the viral DNA replication centers in both OriR-transfected cells and virus-infected cells, highlighting a direct involvement of NP1 in viral DNA replication at OriR. Overall, our study revealed the characteristics of HBoV1 DNA replication at OriR, suggesting novel characteristics of autonomous parvovirus DNA replication. IMPORTANCE Human bocavirus 1 (HBoV1) causes acute respiratory tract infections in young children. The duplex HBoV1 genome replicates in HEK293 cells and produces progeny virions that are infectious in well-differentiated airway epithelial cells. A recombinant AAV2 vector pseudotyped with an HBoV1 capsid has been developed to efficiently deliver the cystic fibrosis transmembrane conductance regulator gene to human airway epithelia. Here, we identified both cis-acting elements and trans-acting proteins that are required for HBoV1 DNA replication at the right-end hairpin in HEK293 cells. We localized the minimal replication origin, which contains both NS1 nicking and binding sites, to a 46-nucleotide sequence in the right-end hairpin. The identification of these essential elements of HBoV1 DNA replication acting both in cis and in trans will provide guidance to develop antiviral strategies targeting viral DNA replication at the right-end hairpin and to design next-generation recombinant HBoV1 vectors, a promising tool for gene therapy of lung diseases. PMID:27334591

  13. Analysis of cis and trans Requirements for DNA Replication at the Right-End Hairpin of the Human Bocavirus 1 Genome.

    PubMed

    Shen, Weiran; Deng, Xuefeng; Zou, Wei; Engelhardt, John F; Yan, Ziying; Qiu, Jianming

    2016-09-01

    Parvoviruses are single-stranded DNA viruses that use the palindromic structures at the ends of the viral genome for their replication. The mechanism of parvovirus replication has been studied mostly in the dependoparvovirus adeno-associated virus 2 (AAV2) and the protoparvovirus minute virus of mice (MVM). Here, we used human bocavirus 1 (HBoV1) to understand the replication mechanism of bocaparvovirus. HBoV1 is pathogenic to humans, causing acute respiratory tract infections, especially in young children under 2 years old. By using the duplex replicative form of the HBoV1 genome in human embryonic kidney 293 (HEK293) cells, we identified the HBoV1 minimal replication origin at the right-end hairpin (OriR). Mutagenesis analyses confirmed the putative NS1 binding and nicking sites within the OriR. Of note, unlike the large nonstructural protein (Rep78/68 or NS1) of other parvoviruses, HBoV1 NS1 did not specifically bind OriR in vitro, indicating that other viral and cellular components or the oligomerization of NS1 is required for NS1 binding to the OriR. In vivo studies demonstrated that residues responsible for NS1 binding and nicking are within the origin-binding domain. Further analysis identified that the small nonstructural protein NP1 is required for HBoV1 DNA replication at OriR. NP1 and other viral nonstructural proteins (NS1 to NS4) colocalized within the viral DNA replication centers in both OriR-transfected cells and virus-infected cells, highlighting a direct involvement of NP1 in viral DNA replication at OriR. Overall, our study revealed the characteristics of HBoV1 DNA replication at OriR, suggesting novel characteristics of autonomous parvovirus DNA replication. Human bocavirus 1 (HBoV1) causes acute respiratory tract infections in young children. The duplex HBoV1 genome replicates in HEK293 cells and produces progeny virions that are infectious in well-differentiated airway epithelial cells. A recombinant AAV2 vector pseudotyped with an HBoV1 capsid has been developed to efficiently deliver the cystic fibrosis transmembrane conductance regulator gene to human airway epithelia. Here, we identified both cis-acting elements and trans-acting proteins that are required for HBoV1 DNA replication at the right-end hairpin in HEK293 cells. We localized the minimal replication origin, which contains both NS1 nicking and binding sites, to a 46-nucleotide sequence in the right-end hairpin. The identification of these essential elements of HBoV1 DNA replication acting both in cis and in trans will provide guidance to develop antiviral strategies targeting viral DNA replication at the right-end hairpin and to design next-generation recombinant HBoV1 vectors, a promising tool for gene therapy of lung diseases. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  14. 32 CFR 293.4 - Definitions.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 2 2014-07-01 2014-07-01 false Definitions. 293.4 Section 293.4 National Defense Department of Defense (Continued) OFFICE OF THE SECRETARY OF DEFENSE (CONTINUED) FREEDOM OF... Definitions. Agency records. (1) A product of data compilation (such as all books, papers, maps, photographs...

  15. 47 CFR 80.293 - Check bearings by authorized ship personnel.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ....293 Section 80.293 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) SAFETY AND SPECIAL... comparison of simultaneous visual and radio direction finder bearings. At least one comparison bearing must... visual bearing relative to the ship's heading and the difference between the visual and radio direction...

  16. STEADY STATE MODELING OF THE MINIMUM CRITICAL CORE OF THE TRANSIENT REACTOR TEST FACILITY

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Anthony L. Alberti; Todd S. Palmer; Javier Ortensi

    2016-05-01

    With the advent of next generation reactor systems and new fuel designs, the U.S. Department of Energy (DOE) has identified the need for the resumption of transient testing of nuclear fuels. The DOE has decided that the Transient Reactor Test Facility (TREAT) at Idaho National Laboratory (INL) is best suited for future testing. TREAT is a thermal neutron spectrum, air-cooled, nuclear test facility that is designed to test nuclear fuels in transient scenarios. These specific scenarios range from simple temperature transients to full fuel melt accidents. DOE has expressed a desire to develop a simulation capability that will accurately modelmore » the experiments before they are irradiated at the facility. It is the aim for this capability to have an emphasis on effective and safe operation while minimizing experimental time and cost. The multi physics platform MOOSE has been selected as the framework for this project. The goals for this work are to identify the fundamental neutronics properties of TREAT and to develop an accurate steady state model for future multiphysics transient simulations. In order to minimize computational cost, the effect of spatial homogenization and angular discretization are investigated. It was found that significant anisotropy is present in TREAT assemblies and to capture this effect, explicit modeling of cooling channels and inter-element gaps is necessary. For this modeling scheme, single element calculations at 293 K gave power distributions with a root mean square difference of 0.076% from those of reference SERPENT calculations. The minimum critical core configuration with identical gap and channel treatment at 293 K resulted in a root mean square, total core, radial power distribution 2.423% different than those of reference SERPENT solutions.« less

  17. Characteristics of thermally-induced transverse cracks in graphite epoxy composite laminates

    NASA Technical Reports Server (NTRS)

    Adams, D. S.; Bowles, D. E.; Herakovich, C. T.

    1983-01-01

    The characteristics of thermally induced transverse cracks in T300/5208 graphite-epoxy cross-ply and quasi-isotropic laminates were investigated both experimentally and analytically. The formation of transverse cracks and the subsequent crack spacing present during cool down to -250 F (116K) and thermal cycling between 250 and -250 F (116 and 394K) was investigated. The state of stress in the vicinity of a transverse crack and the influence of transverse cracking on the laminate coefficient of thermal expansion (CTE) was predicted using a generalized plane strain finite element analysis and a modified shear lag analysis. A majority of the cross-ply laminates experienced transverse cracking during the initial cool down to -250 F whereas the quasi-isotropic laminates remained uncracked. The in situ transverse strength of the 90 degree layers was more than 1.9 times greater than the transverse strength of the unidirectional 90 degree material for all laminates investigated.

  18. 10 CFR 431.293 - Materials incorporated by reference.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    .... Department of Energy, Office of Energy Efficiency and Renewable Energy, Building Technologies Program, 6th... 10 Energy 3 2010-01-01 2010-01-01 false Materials incorporated by reference. 431.293 Section 431.293 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY PROGRAM FOR CERTAIN COMMERCIAL...

  19. 10 CFR 431.293 - Materials incorporated by reference.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    .... Department of Energy, Office of Energy Efficiency and Renewable Energy, Building Technologies Program, 6th... 10 Energy 3 2011-01-01 2011-01-01 false Materials incorporated by reference. 431.293 Section 431.293 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY PROGRAM FOR CERTAIN COMMERCIAL...

  20. 10 CFR 431.293 - Materials incorporated by reference.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    .... Department of Energy, Office of Energy Efficiency and Renewable Energy, Building Technologies Program, 6th... 10 Energy 3 2013-01-01 2013-01-01 false Materials incorporated by reference. 431.293 Section 431.293 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY PROGRAM FOR CERTAIN COMMERCIAL...

  1. 10 CFR 431.293 - Materials incorporated by reference.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    .... Department of Energy, Office of Energy Efficiency and Renewable Energy, Building Technologies Program, 6th... 10 Energy 3 2014-01-01 2014-01-01 false Materials incorporated by reference. 431.293 Section 431.293 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY PROGRAM FOR CERTAIN COMMERCIAL...

  2. 10 CFR 431.293 - Materials incorporated by reference.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    .... Department of Energy, Office of Energy Efficiency and Renewable Energy, Building Technologies Program, 6th... 10 Energy 3 2012-01-01 2012-01-01 false Materials incorporated by reference. 431.293 Section 431.293 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY PROGRAM FOR CERTAIN COMMERCIAL...

  3. 28 CFR 2.93 - Travel approval.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Travel approval. 2.93 Section 2.93... Travel approval. (a) A parolee's Supervision Officer may approve travel outside the district of... possibilities. (3) Recurring travel across a district boundary, not to exceed fifty miles outside the district...

  4. 14 CFR 293.10 - Exemption.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Exemption. 293.10 Section 293.10 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION (AVIATION PROCEEDINGS) ECONOMIC... exempted from the duty to file all passenger tariffs except those setting forth one-way economy-class fares...

  5. 36 CFR 293.6 - Commercial enterprises, roads, motor vehicles, motorized equipment, motorboats, aircraft...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., structures, and cutting of trees. 293.6 Section 293.6 Parks, Forests, and Public Property FOREST SERVICE... trees. Except as provided in the Wilderness Act, subsequent legislation establishing a particular... from aircraft; no structures or installations; and no cutting of trees for nonwilderness purposes. (a...

  6. 36 CFR 293.6 - Commercial enterprises, roads, motor vehicles, motorized equipment, motorboats, aircraft...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ..., structures, and cutting of trees. 293.6 Section 293.6 Parks, Forests, and Public Property FOREST SERVICE... trees. Except as provided in the Wilderness Act, subsequent legislation establishing a particular... from aircraft; no structures or installations; and no cutting of trees for nonwilderness purposes. (a...

  7. 36 CFR 293.6 - Commercial enterprises, roads, motor vehicles, motorized equipment, motorboats, aircraft...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., structures, and cutting of trees. 293.6 Section 293.6 Parks, Forests, and Public Property FOREST SERVICE... trees. Except as provided in the Wilderness Act, subsequent legislation establishing a particular... from aircraft; no structures or installations; and no cutting of trees for nonwilderness purposes. (a...

  8. 36 CFR 293.6 - Commercial enterprises, roads, motor vehicles, motorized equipment, motorboats, aircraft...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ..., structures, and cutting of trees. 293.6 Section 293.6 Parks, Forests, and Public Property FOREST SERVICE... trees. Except as provided in the Wilderness Act, subsequent legislation establishing a particular... from aircraft; no structures or installations; and no cutting of trees for nonwilderness purposes. (a...

  9. 5 CFR 293.305 - Type of folder to be used.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Type of folder to be used. 293.305 Section 293.305 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS... OPFs from Office of Federal Supply and Services stock (Standard Form 66) for the folders required by...

  10. 5 CFR 293.202 - Records subject to Office or agency Privacy Act regulations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Privacy Act regulations. 293.202 Section 293.202 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT... Records subject to Office or agency Privacy Act regulations. When the Office of Personnel Management publishes in the Federal Register a notice of system of records for personnel records which are maintained...

  11. 5 CFR 293.309 - Reconstruction of lost OPFs.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Reconstruction of lost OPFs. 293.309 Section 293.309 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS... necessary precautions to safeguard all OPFs. In the event of a lost or destroyed OPF, the current (or last...

  12. 36 CFR 293.3 - Control of uses.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 2 2010-07-01 2010-07-01 false Control of uses. 293.3 Section 293.3 Parks, Forests, and Public Property FOREST SERVICE, DEPARTMENT OF AGRICULTURE WILDERNESS... legislation establishing a particular unit, or the regulations in this part, the Chief, Forest Service, may...

  13. 5 CFR 293.309 - Reconstruction of lost OPFs.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Reconstruction of lost OPFs. 293.309 Section 293.309 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS... necessary precautions to safeguard all OPFs. In the event of a lost or destroyed OPF, the current (or last...

  14. 36 CFR 293.7 - Grazing of livestock.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 2 2010-07-01 2010-07-01 false Grazing of livestock. 293.7...-PRIMITIVE AREAS § 293.7 Grazing of livestock. (a) The grazing of livestock, where such use was established..., shall be permitted to continue under the general regulations covering grazing of livestock on the...

  15. 36 CFR 293.3 - Control of uses.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 2 2011-07-01 2011-07-01 false Control of uses. 293.3 Section 293.3 Parks, Forests, and Public Property FOREST SERVICE, DEPARTMENT OF AGRICULTURE WILDERNESS... legislation establishing a particular unit, or the regulations in this part, the Chief, Forest Service, may...

  16. KSC-06pd2382

    NASA Image and Video Library

    2006-07-05

    JOHNSON SPACE CENTER, Houston, Texas -- STS116-S-001 (July 2006) - The STS-116 patch design signifies the continuing assembly of the International Space Station (ISS). The primary mission objective is to deliver and install the P5 truss element. The P5 installation will be conducted during the first of three planned spacewalks, and will involve use of both the shuttle and station robotic arms. The remainder of the mission will include a major reconfiguration and activation of the ISS electrical and thermal control systems, as well as delivery of Zvezda Service Module debris panels, which will increase ISS protection from potential impacts of micro-meteorites and orbital debris. In addition, a single expedition crew member will launch on STS-116 to remain onboard the station, replacing an expedition crew member who will fly home with the shuttle crew. The crew patch depicts the space shuttle rising above the Earth and ISS. The United States and Swedish flags trail the orbiter, depicting the international composition of the STS-116 crew. The seven stars of the constellation Ursa Major are used to provide direction to the North Star, which is superimposed over the installation location of the P5 truss on ISS. The NASA insignia design for space shuttle space flights is reserved for use by the astronauts and other official use as the NASA Administrator may authorize. Public availability has been approved only in the form of illustrations by the various news media. When and if there is any change in this policy, which is not anticipated, such will be publicly announced.

  17. X-ray fluorescence microscopy artefacts in elemental maps of topologically complex samples: Analytical observations, simulation and a map correction method

    NASA Astrophysics Data System (ADS)

    Billè, Fulvio; Kourousias, George; Luchinat, Enrico; Kiskinova, Maya; Gianoncelli, Alessandra

    2016-08-01

    XRF spectroscopy is among the most widely used non-destructive techniques for elemental analysis. Despite the known angular dependence of X-ray fluorescence (XRF), topological artefacts remain an unresolved issue when using X-ray micro- or nano-probes. In this work we investigate the origin of the artefacts in XRF imaging of topologically complex samples, which are unresolved problems in studies of organic matter due to the limited travel distances of low energy XRF emission from the light elements. In particular we mapped Human Embryonic Kidney (HEK293T) cells. The exemplary results with biological samples, obtained with a soft X-ray scanning microscope installed at a synchrotron facility were used for testing a mathematical model based on detector response simulations, and for proposing an artefact correction method based on directional derivatives. Despite the peculiar and specific application, the methodology can be easily extended to hard X-rays and to set-ups with multi-array detector systems when the dimensions of surface reliefs are in the order of the probing beam size.

  18. Cell Context Dependent p53 Genome-Wide Binding Patterns and Enrichment at Repeats

    DOE PAGES

    Botcheva, Krassimira; McCorkle, Sean R.

    2014-11-21

    The p53 ability to elicit stress specific and cell type specific responses is well recognized, but how that specificity is established remains to be defined. Whether upon activation p53 binds to its genomic targets in a cell type and stress type dependent manner is still an open question. Here we show that the p53 binding to the human genome is selective and cell context-dependent. We mapped the genomic binding sites for the endogenous wild type p53 protein in the human cancer cell line HCT116 and compared them to those we previously determined in the normal cell line IMR90. We reportmore » distinct p53 genome-wide binding landscapes in two different cell lines, analyzed under the same treatment and experimental conditions, using the same ChIP-seq approach. This is evidence for cell context dependent p53 genomic binding. The observed differences affect the p53 binding sites distribution with respect to major genomic and epigenomic elements (promoter regions, CpG islands and repeats). We correlated the high-confidence p53 ChIP-seq peaks positions with the annotated human repeats (UCSC Human Genome Browser) and observed both common and cell line specific trends. In HCT116, the p53 binding was specifically enriched at LINE repeats, compared to IMR90 cells. The p53 genome-wide binding patterns in HCT116 and IMR90 likely reflect the different epigenetic landscapes in these two cell lines, resulting from cancer-associated changes (accumulated in HCT116) superimposed on tissue specific differences (HCT116 has epithelial, while IMR90 has mesenchymal origin). In conclusion, our data support the model for p53 binding to the human genome in a highly selective manner, mobilizing distinct sets of genes, contributing to distinct pathways.« less

  19. Finite element simulations of the Portevin Le Chatelier effect in aluminium alloy

    NASA Astrophysics Data System (ADS)

    Hopperstad, O. S.; Børvik, T.; Berstad, T.; Benallal, A.

    2006-08-01

    Finite element simulations of the Portevin-Le Chatelier effect in aluminium alloy 5083-H116 are presented and evaluated against existing experimental results. The constitutive model of McCormick (1988) for materials exhibiting negative steady-state strain-rate sensitivity is incorporated into an elastic-viscoplastic model for large plastic deformations and implemented in LS-DYNA for use with the explicit or implicit solver. Axisymmetric tensile specimens loaded at different strain rates are studied numerically, and it is shown that the model predicts the experimental behaviour with reasonable accuracy; including serrated yielding and propagating bands of localized plastic deformation along the gauge length of the specimen at intermediate strain rates.

  20. Retroviral vectors encoding ADA regulatory locus control region provide enhanced T-cell-specific transgene expression.

    PubMed

    Trinh, Alice T; Ball, Bret G; Weber, Erin; Gallaher, Timothy K; Gluzman-Poltorak, Zoya; Anderson, French; Basile, Lena A

    2009-12-30

    Murine retroviral vectors have been used in several hundred gene therapy clinical trials, but have fallen out of favor for a number of reasons. One issue is that gene expression from viral or internal promoters is highly variable and essentially unregulated. Moreover, with retroviral vectors, gene expression is usually silenced over time. Mammalian genes, in contrast, are characterized by highly regulated, precise levels of expression in both a temporal and a cell-specific manner. To ascertain if recapitulation of endogenous adenosine deaminase (ADA) expression can be achieved in a vector construct we created a new series of Moloney murine leukemia virus (MuLV) based retroviral vector that carry human regulatory elements including combinations of the ADA promoter, the ADA locus control region (LCR), ADA introns and human polyadenylation sequences in a self-inactivating vector backbone. A MuLV-based retroviral vector with a self-inactivating (SIN) backbone, the phosphoglycerate kinase promoter (PGK) and the enhanced green fluorescent protein (eGFP), as a reporter gene, was generated. Subsequent vectors were constructed from this basic vector by deletion or addition of certain elements. The added elements that were assessed are the human ADA promoter, human ADA locus control region (LCR), introns 7, 8, and 11 from the human ADA gene, and human growth hormone polyadenylation signal. Retroviral vector particles were produced by transient three-plasmid transfection of 293T cells. Retroviral vectors encoding eGFP were titered by transducing 293A cells, and then the proportion of GFP-positive cells was determined using fluorescence-activated cell sorting (FACS). Non T-cell and T-cell lines were transduced at a multiplicity of infection (MOI) of 0.1 and the yield of eGFP transgene expression was evaluated by FACS analysis using mean fluorescent intensity (MFI) detection. Vectors that contained the ADA LCR were preferentially expressed in T-cell lines. Further improvements in T-cell specific gene expression were observed with the incorporation of additional cis-regulatory elements, such as a human polyadenylation signal and intron 7 from the human ADA gene. These studies suggest that the combination of an authentically regulated ADA gene in a murine retroviral vector, together with additional locus-specific regulatory refinements, will yield a vector with a safer profile and greater efficacy in terms of high-level, therapeutic, regulated gene expression for the treatment of ADA-deficient severe combined immunodeficiency.

  1. Inhibition of NAMPT pathway by FK866 activates the function of p53 in HEK293T cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Thakur, Basant Kumar, E-mail: thakur.basant@mh-hannover.de; Department of Molecular Hematopoiesis, Hannover Medical School, Carl Neuberg Str-1, 30625 Hannover; Dittrich, Tino

    2012-08-03

    Highlights: Black-Right-Pointing-Pointer In 293T cells, p53 is considered to be inactive due to its interaction with the large T-antigen. Black-Right-Pointing-Pointer Acetylation of p53 at lysine 382 is important for its functional activation. Black-Right-Pointing-Pointer First evidence to document the presence of a functional p53 in 293T cells. Black-Right-Pointing-Pointer Inhibition of NAMPT/SIRT pathway by FK866 in 293T cells increases the functional activity of p53. Black-Right-Pointing-Pointer This activation of p53 involves reversible acetylation of p53 at lysine 382. -- Abstract: Inactivation of p53 protein by endogenous and exogenous carcinogens is involved in the pathogenesis of different human malignancies. In cancer associated with SV-40more » DNA tumor virus, p53 is considered to be non-functional mainly due to its interaction with the large T-antigen. Using the 293T cell line (HEK293 cells transformed with large T antigen) as a model, we provide evidence that p53 is one of the critical downstream targets involved in FK866-mediated killing of 293T cells. A reduced rate of apoptosis and an increased number of cells in S-phase was accompanied after knockdown of p53 in these cells. Inhibition of NAMPT by FK866, or inhibition of SIRT by nicotinamide decreased proliferation and triggered death of 293T cells involving the p53 acetylation pathway. Additionally, knockdown of p53 attenuated the effect of FK866 on cell proliferation, apoptosis, and cell cycle arrest. The data presented here shed light on two important facts: (1) that p53 in 293T cells is active in the presence of FK866, an inhibitor of NAMPT pathway; (2) the apoptosis induced by FK866 in 293T cells is associated with increased acetylation of p53 at Lys382, which is required for the functional activity of p53.« less

  2. Research misconduct oversight: defining case costs.

    PubMed

    Gammon, Elizabeth; Franzini, Luisa

    2013-01-01

    This study uses a sequential mixed method study design to define cost elements of research misconduct among faculty at academic medical centers. Using time driven activity based costing, the model estimates a per case cost for 17 cases of research misconduct reported by the Office of Research Integrity for the period of 2000-2005. Per case cost of research misconduct was found to range from $116,160 to $2,192,620. Research misconduct cost drivers are identified.

  3. Crocin and quercetin prevent PAT-induced apoptosis in mammalian cells: Involvement of ROS-mediated ER stress pathway.

    PubMed

    Boussabbeh, Manel; Prola, Alexandre; Ben Salem, Intidhar; Guilbert, Arnaud; Bacha, Hassen; Lemaire, Christophe; Abis-Essefi, Salwa

    2016-12-01

    Patulin (PAT) is a secondary metabolite produced by several species of the genera of Penicillium, Aspergillus, and Byssochlamys that can be found in rotting fruits, especially in apples and apple-based products. Exposure to this mycotoxin has been reported to induce intestinal and kidney injuries. The mechanism underlying such toxicity has been linked to the induction of apoptosis which occurred with reactive oxygen species production and endoplasmic reticulum (ER) stress induction. This study aimed to evaluate the effect of the two common dietary compounds Quercetin (QUER), a natural flavonoid, and Crocin (CRO), a natural carotenoid, on PAT-induced toxicity in human colon carcinoma (HCT116) and embryonic kidney cells (HEK293). We showed that antioxidant properties of QUER and CRO help to prevent ER stress activation and lipid peroxidation as evidenced by the reduction in GRP78 and GADD34 expressions and the decrease in malondialdehyde production. Furthermore, we demonstrated their ability to re-establish the loss of the mitochondrial membrane potential to inhibit caspase 3 activation and DNA fragmentation. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1851-1858, 2016. © 2015 Wiley Periodicals, Inc.

  4. Dual-targeting siRNAs

    PubMed Central

    Tiemann, Katrin; Höhn, Britta; Ehsani, Ali; Forman, Stephen J.; Rossi, John J.; Sætrom, Pål

    2010-01-01

    We have developed an algorithm for the prediction of dual-targeting short interfering RNAs (siRNAs) in which both strands are deliberately designed to separately target different mRNA transcripts with complete complementarity. An advantage of this approach versus the use of two separate duplexes is that only two strands, as opposed to four, are competing for entry into the RNA-induced silencing complex. We chose to design our dual-targeting siRNAs as Dicer substrate 25/27mer siRNAs, since design features resembling pre-microRNAs (miRNAs) can be introduced for Dicer processing. Seven different dual-targeting siRNAs targeting genes that are potential targets in cancer therapy have been developed including Bcl2, Stat3, CCND1, BIRC5, and MYC. The dual-targeting siRNAs have been characterized for dual target knockdown in three different cell lines (HEK293, HCT116, and PC3), where they were as effective as their corresponding single-targeting siRNAs in target knockdown. The algorithm developed in this study should prove to be useful for predicting dual-targeting siRNAs in a variety of different targets and is available from http://demo1.interagon.com/DualTargeting/. PMID:20410240

  5. 36 CFR 293.16 - Special provisions governing the Boundary Waters Canoe Area Wilderness, Superior National Forest...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 36 Parks, Forests, and Public Property 2 2012-07-01 2012-07-01 false Special provisions governing the Boundary Waters Canoe Area Wilderness, Superior National Forest, Minnesota. 293.16 Section 293.16... Forest, Minnesota. (a) Motorboat use. (1) For purposes of this section, motorboats permitted to operate...

  6. 36 CFR 293.16 - Special provisions governing the Boundary Waters Canoe Area Wilderness, Superior National Forest...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 36 Parks, Forests, and Public Property 2 2013-07-01 2013-07-01 false Special provisions governing the Boundary Waters Canoe Area Wilderness, Superior National Forest, Minnesota. 293.16 Section 293.16... Forest, Minnesota. (a) Motorboat use. (1) For purposes of this section, motorboats permitted to operate...

  7. 36 CFR 293.16 - Special provisions governing the Boundary Waters Canoe Area Wilderness, Superior National Forest...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 36 Parks, Forests, and Public Property 2 2014-07-01 2014-07-01 false Special provisions governing the Boundary Waters Canoe Area Wilderness, Superior National Forest, Minnesota. 293.16 Section 293.16... Forest, Minnesota. (a) Motorboat use. (1) For purposes of this section, motorboats permitted to operate...

  8. 36 CFR 293.6 - Commercial enterprises, roads, motor vehicles, motorized equipment, motorboats, aircraft...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 2 2011-07-01 2011-07-01 false Commercial enterprises, roads..., structures, and cutting of trees. 293.6 Section 293.6 Parks, Forests, and Public Property FOREST SERVICE... forms of mechanical transport; no landing of aircraft; no dropping of materials, supplies, or persons...

  9. 76 FR 67418 - Request for Comments on NIST Special Publication 500-293, US Government Cloud Computing...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-01

    ...-1659-01] Request for Comments on NIST Special Publication 500-293, US Government Cloud Computing... Publication 500-293, US Government Cloud Computing Technology Roadmap, Release 1.0 (Draft). This document is... (USG) agencies to accelerate their adoption of cloud computing. The roadmap has been developed through...

  10. 36 CFR 293.11 - Water rights.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 2 2010-07-01 2010-07-01 false Water rights. 293.11 Section...-PRIMITIVE AREAS § 293.11 Water rights. Nothing in the regulations in this part constitutes an expressed or implied claim or denial on the part of the Department of Agriculture as to exemption from State water laws. ...

  11. 36 CFR 293.11 - Water rights.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 2 2011-07-01 2011-07-01 false Water rights. 293.11 Section...-PRIMITIVE AREAS § 293.11 Water rights. Nothing in the regulations in this part constitutes an expressed or implied claim or denial on the part of the Department of Agriculture as to exemption from State water laws. ...

  12. 36 CFR 293.11 - Water rights.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 36 Parks, Forests, and Public Property 2 2013-07-01 2013-07-01 false Water rights. 293.11 Section...-PRIMITIVE AREAS § 293.11 Water rights. Nothing in the regulations in this part constitutes an expressed or implied claim or denial on the part of the Department of Agriculture as to exemption from State water laws. ...

  13. 36 CFR 293.11 - Water rights.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 36 Parks, Forests, and Public Property 2 2012-07-01 2012-07-01 false Water rights. 293.11 Section...-PRIMITIVE AREAS § 293.11 Water rights. Nothing in the regulations in this part constitutes an expressed or implied claim or denial on the part of the Department of Agriculture as to exemption from State water laws. ...

  14. 36 CFR 293.11 - Water rights.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 36 Parks, Forests, and Public Property 2 2014-07-01 2014-07-01 false Water rights. 293.11 Section...-PRIMITIVE AREAS § 293.11 Water rights. Nothing in the regulations in this part constitutes an expressed or implied claim or denial on the part of the Department of Agriculture as to exemption from State water laws. ...

  15. 5 CFR 293.405 - Disposition of records.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... RECORDS Employee Performance File System Records § 293.405 Disposition of records. (a) When the OPF of a..., employee performance files shall be forwarded to gaining agencies at the same time as the OPF (5 CFR 293... former employee returns to an agency, a new employee performance file will be created unless the prior...

  16. 5 CFR 293.405 - Disposition of records.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... RECORDS Employee Performance File System Records § 293.405 Disposition of records. (a) When the OPF of a..., employee performance files shall be forwarded to gaining agencies at the same time as the OPF (5 CFR 293... former employee returns to an agency, a new employee performance file will be created unless the prior...

  17. 5 CFR 293.405 - Disposition of records.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... RECORDS Employee Performance File System Records § 293.405 Disposition of records. (a) When the OPF of a..., employee performance files shall be forwarded to gaining agencies at the same time as the OPF (5 CFR 293... former employee returns to an agency, a new employee performance file will be created unless the prior...

  18. 5 CFR 293.405 - Disposition of records.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... RECORDS Employee Performance File System Records § 293.405 Disposition of records. (a) When the OPF of a..., employee performance files shall be forwarded to gaining agencies at the same time as the OPF (5 CFR 293... former employee returns to an agency, a new employee performance file will be created unless the prior...

  19. 5 CFR 293.405 - Disposition of records.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... RECORDS Employee Performance File System Records § 293.405 Disposition of records. (a) When the OPF of a..., employee performance files shall be forwarded to gaining agencies at the same time as the OPF (5 CFR 293... former employee returns to an agency, a new employee performance file will be created unless the prior...

  20. Molecular Imaging of Phosphorylation Events for Drug Development

    PubMed Central

    Chan, C. T.; Paulmurugan, R.; Reeves, R. E.; Solow-Cordero, D.; Gambhir, S. S.

    2014-01-01

    Purpose Protein phosphorylation mediated by protein kinases controls numerous cellular processes. A genetically encoded, generalizable split firefly luciferase (FL)-assisted complementation system was developed for noninvasive monitoring phosphorylation events and efficacies of kinase inhibitors in cell culture and in small living subjects by optical bioluminescence imaging. Procedures An Akt sensor (AST) was constructed to monitor Akt phosphorylation and the effect of different PI-3K and Akt inhibitors. Specificity of AST was determined using a non-phosphorylable mutant sensor containing an alanine substitution (ASA). Results The PI-3K inhibitor LY294002 and Akt kinase inhibitor perifosine led to temporal- and dose-dependent increases in complemented FL activities in 293T human kidney cancer cells stably expressing AST (293T/AST) but not in 293T/ASA cells. Inhibition of endogenous Akt phosphorylation and kinase activities by perifosine also correlated with increase in complemented FL activities in 293T/AST cells but not in 293T/ASA cells. Treatment of nude mice bearing 293T/AST xenografts with perifosine led to a 2-fold increase in complemented FL activities compared to that of 293T/ASA xenografts. Our system was used to screen a small chemical library for novel modulators of Akt kinase activity. Conclusion This generalizable approach for noninvasive monitoring of phosphorylation events will accelerate the discovery and validation of novel kinase inhibitors and modulators of phosphorylation events. PMID:19048345

  1. Comparison of the Thermal Expansion Behavior of Several Intermetallic Silicide Alloys Between 293 and 1523 K

    NASA Technical Reports Server (NTRS)

    Raj, Sai V.

    2014-01-01

    Thermal expansion measurements were conducted on hot-pressed CrSi(sub 2), TiSi(sub 2), W Si(sub 2) and a two-phase Cr-Mo-Si intermetallic alloy between 293 and 1523 K during three heat-cool cycles. The corrected thermal expansion, (L/L(sub 0)(sub thermal), varied with the absolute temperature, T, as (deltaL/L(sub 0)(sub thermal) = A(T-293)(sup 3) + B(T-293)(sup 2) + C(T-293) + D, where A, B, C and D are regression constants. Excellent reproducibility was observed for most of the materials after the first heat-up cycle. In some cases, the data from the first heatup cycle deviated from those determined in the subsequent cycles. This deviation was attributed to the presence of residual stresses developed during processing, which are relieved after the first heat-up cycle.

  2. [Novel bidirectional promoter from human genome].

    PubMed

    Orekhova, A S; Sverdlova, P S; Spirin, P V; Leonova, O G; Popenko, V I; Prasolov, V S; Rubtsov, P M

    2011-01-01

    In human and other mammalian genomes a number of closely linked gene pairs transcribed in opposite directions are found. According to bioinformatic analysis up to 10% of human genes are arranged in this way. In present work the fragment of human genome was cloned that separates genes localized at 2p13.1 and oriented "head-to-head", coding for hypothetical proteins with unknown functions--CCDC (Coiled Coil Domain Containing) 142 and TTC (TetraTricopeptide repeat Containing) 31. Intergenic CCDC142-TTC31 region overlaps with CpG-island and contains a number of potential binding sites for transcription factors. This fragment functions as bidirectional promoter in the system ofluciferase reporter gene expression upon transfection of human embryonic kidney (HEK293) cells. The vectors containing genes of two fluorescent proteins--green (EGFP) and red (DsRed2) in opposite orientations separated by the fragment of CCDC142-TTC31 intergenic region were constructed. In HEK293 cells transfected with these vectors simultaneous expression of two fluorescent proteins is observed. Truncated versions of intergenic region were obtained and their promoter activity measured. Minimal promoter fragment contains elements Inr, BRE, DPE characteristic for TATA-less promoters. Thus, from the human genome the novel bidirectional promoter was cloned that can be used for simultaneous constitutive expression of two genes in human cells.

  3. Reading Level and Length of Written Research Consent Forms

    PubMed Central

    Foe, Gabriella; Lally, Rachel

    2015-01-01

    Abstract In 100 Institutional Review Board approved consent forms (CFs), we assessed pages, reading levels, and whether they included essential elements. CF page numbers ranged from 3 to 28 (mean, 10.3) and readability ranged from grades 5.6 to 28.9 (mean, 11.6). The CF mean score for including essential elements was 90.8% (range: 63.5–100%). There were no significant differences by specialty in number of pages (p = 0.053), but surgical specialties had the highest readability (mean, 13.1), and pediatrics the lowest (10.5), p = 0.008. While approved CFs generally included the Office for Human Research Protections recommended essential elements, they were very long, and even pediatric forms, which had the lowest reading levels, were written on average at a tenth grade level. Researchers need guidance to resolve pressure between regulatory mandates and guidelines and “keeping it simple and clear.” PMID:25580939

  4. Buckling Testing and Analysis of Honeycomb Sandwich Panel Arc Segments of a Full-Scale Fairing Barrel: Comparison of In- and Out-of-Autoclave Facesheet Configurations

    NASA Technical Reports Server (NTRS)

    Pineda, Evan Jorge; Myers, David E.; Kosareo, Daniel N.; Zalewski, Bart F.; Kellas, Sotiris; Dixon, Genevieve D.; Krivanek, Thomas M.; Gyekenyesi, Thomas G.

    2014-01-01

    Four honeycomb sandwich panels, representing 1/16th arc segments of a 10-m diameter barrel section of the Heavy Lift Launch Vehicle, were manufactured and tested under the NASA Composites for Exploration and the NASA Constellation Ares V programs. Two configurations were chosen for the panels: 6-ply facesheets with 1.125 in. honeycomb core and 8-ply facesheets with 1.0 in. honeycomb core. Additionally, two separate carbon fiber/epoxy material systems were chosen for the facesheets: in-autoclave IM7/977-3 and out-of-autoclave T40-800b/5320-1. Smaller 3 ft. by 5 ft. panels were cut from the 1/16th barrel sections and tested under compressive loading. Furthermore, linear eigenvalue and geometrically nonlinear finite element analyses were performed to predict the compressive response of each 3 ft. by 5 ft. panel. To improve the robustness of the geometrically nonlinear finite element model, measured surface imperfections were included in the geometry of the model. Both the linear and nonlinear models yielded good qualitative and quantitative predictions. Additionally, it was correctly predicted that the panel would fail in buckling prior to failing in strength. Furthermore, several imperfection studies were performed to investigate the influence of geometric imperfections, fiber angle misalignments, and three-dimensional effects on the compressive response of the panel.

  5. Chorein interacts with α-tubulin and histone deacetylase 6, and overexpression preserves cell viability during nutrient deprivation in human embryonic kidney 293 cells.

    PubMed

    Sasaki, Natsuki; Nakamura, Masayuki; Kodama, Akiko; Urata, Yuka; Shiokawa, Nari; Hayashi, Takehiro; Sano, Akira

    2016-11-01

    The autophagy pathway has recently been implicated in several neurodegenerative diseases. Recently, it was reported that chorein-depleted cells showed accumulation of autophagic markers and impaired autophagic flux. Here, we demonstrate that chorein overexpression preserves cell viability from starvation-induced cell death in human embryonic kidney 293 (HEK293) cells. Subsequent coimmunoprecipitation and reverse coimmunoprecipitation assays using extracts from chorein that stably overexpressed HEK293 cells revealed that chorein interacts with α-tubulin and histone deacetylase 6, a known α-tubulin deacetylater and central component of basal autophagy. Indeed, acetylated α-tubulin immunoreactivity was significantly decreased in chorein that stably overexpressed HEK293 cells. These results suggest that chorein/histone deacetylase 6/α-tubulin interactions may play an important role in starvation-induced cell stress, and their disruption may be one of the molecular pathogenic mechanisms of chorea-acanthocytosis.-Sasaki, N., Nakamura, M., Kodama, A., Urata, Y., Shiokawa, N., Hayashi, T., Sano, A. Chorein interacts with α-tubulin and histone deacetylase 6, and overexpression preserves cell viability during nutrient deprivation in human embryonic kidney 293 cells. © FASEB.

  6. The genome-wide expression profile of Curcuma longa-treated cisplatin-stimulated HEK293 cells

    PubMed Central

    Sohn, Sung-Hwa; Ko, Eunjung; Chung, Hwan-Suck; Lee, Eun-Young; Kim, Sung-Hoon; Shin, Minkyu; Hong, Moochang; Bae, Hyunsu

    2010-01-01

    AIM The rhizome of turmeric, Curcuma longa (CL), is a herbal medicine used in many traditional prescriptions. It has previously been shown that CL treatment showed greater than 47% recovery from cisplatin-induced cell damage in human kidney HEK 293 cells. This study was conducted to evaluate the recovery mechanisms of CL that occur during cisplatin induced nephrotoxicity by examining the genome wide mRNA expression profiles of HEK 293 -cells. METHOD Recovery mechanisms of CL that occur during cisplatin-induced nephrotoxicity were determined by microarray, real-time PCR, immunofluorescent confocal microscopy and Western blot analysis. RESULTS The results of microarray analysis and real-time PCR revealed that NFκB pathway-related genes and apoptosis-related genes were down-regulated in CL-treated HEK 293 cells. In addition, immunofluorescent confocal microscopy and Western blot analysis revealed that NFκB p65 nuclear translocation was inhibited in CL-treated HEK 293 cells. Therefore, the mechanism responsible for the effects of CL on HEK 293 cells is closely associated with regulation of the NFκB pathway. CONCLUSION CL possesses novel therapeutic agents that can be used for the prevention or treatment of cisplatin-induced renal disorders. PMID:20840446

  7. Rational modification of Corynebacterium glutamicum dihydrodipicolinate reductase to switch the nucleotide-cofactor specificity for increasing l-lysine production.

    PubMed

    Xu, Jian-Zhong; Yang, Han-Kun; Liu, Li-Ming; Wang, Ying-Yu; Zhang, Wei-Guo

    2018-03-25

    l-lysine is an important amino acid in animals and humans and NADPH is a vital cofactor for maximizing the efficiency of l-lysine fermentation. Dihydrodipicolinate reductase (DHDPR), an NAD(P)H-dependent enzyme, shows a variance in nucleotide-cofactor affinity in bacteria. In this study, we rationally engineered Corynebacterium glutamicum DHDPR (CgDHDPR) to switch its nucleotide-cofactor specificity resulting in an increase in final titer (from 82.6 to 117.3 g L -1 ), carbon yield (from 0.35 to 0.44 g [g glucose] -1 ) and productivity (from 2.07 to 2.93 g L -1  hr -1 ) of l-lysine in JL-6 ΔdapB::Ec-dapB C115G,G116C in fed-batch fermentation. To do this, we comparatively analyzed the characteristics of CgDHDPR and Escherichia coli DHDPR (EcDHDPR), indicating that hetero-expression of NADH-dependent EcDHDPR increased l-lysine production. Subsequently, we rationally modified the conserved structure of cofactor-binding motif, and results indicated that introducing the mutation K11A or R13A in CgDHDPR and introducing the mutation R16A or R39A in EcDHDPR modifies the nucleotide-cofactor affinity of DHDPR. Lastly, the effects of these mutated DHDPRs on l-lysine production were investigated. The highest increase (26.2%) in l-lysine production was observed for JL-6 ΔdapB::Ec-dapB C115G,G116C , followed by JL-6 Cg-dapB C37G,G38C (21.4%) and JL-6 ΔdapB::Ec-dapB C46G,G47C (15.2%). This is the first report of a rational modification of DHDPR that enhances the l-lysine production and yield through the modulation of nucleotide-cofactor specificity. © 2018 Wiley Periodicals, Inc.

  8. [Not Available].

    PubMed

    Angulo, Daniela; Bustos, Edson; Sánchez, Andrés; Barja, Salesa

    2016-07-19

    Introducción: la rehabilitación de la alimentación por vía oral (RVO) es compleja en pacientes que han recibido nutrición enteral (NE) prolongada. Objetivo: describir este proceso en niños con enfermedades respiratorias crónicas y sonda nasoenteral (SNE) o gastrostomía (GT).Pacientes y métodos: estudio retrospectivo con revisión de registros clínicos de niños con NE mayor a dos meses, ingresados entre 2005 y 2014 al Hospital Josefina Martínez.Resultados: Se incluyeron 116 pacientes, con mediana de edad 10 meses (Rango: 3 a 101), 56% hombres. Diagnóstico: 34,5% Daño pulmonar crónico postinfeccioso (DPC), 29,3% Insuficiencia respiratoria por enfermedad neuromuscular, 19% Displasia broncopulmonar y 17,2% enfermedad de la vía aérea. Con traqueostomía: 82,8%. Eran usuarios de GT 89,7% y de SNG 10,3%, instaladas con mediana de edad 6 meses (0 a 74), por ingesta insuficiente (6,6%) o trastorno de deglución (92,4%). Del grupo total, 36,2% (42/116) tenía indicación de RVO, los cuales habían recibido NE durante 12,2 meses (2 a 41); de estos 50% (21/42) logró alimentarse exclusivamente por vía oral (91% SNG y 35,4% GT, Chi2 p = 0,023), 14% parcialmente y 36% no lo logró. El tiempo para lograr la vía oral exclusiva fue de 9,75 meses (0,5 a 47), sin diferencia por edad, sexo, vía de acceso, duración NE ni presencia de enfermedad neurológica.Conclusión: en pacientes con enfermedades respiratorias crónicas graves y NE prolongada, la RVO es un proceso lento pero posible: 64% lo logra de modo completo o parcial.

  9. Multielement analysis of interplanetary dust particles using TOF-SIMS

    NASA Technical Reports Server (NTRS)

    Stephan, T.; Kloeck, W.; Jessberger, E. K.; Rulle, H.; Zehnpfenning, J.

    1993-01-01

    Sections of three stratospheric particles (U2015G1, W7029*A27, and L2005P9) were analyzed with TOF-SIMS (Time Of Flight-Secondary Ion Mass Spectrometry) continuing our efforts to investigate the element distribution in interplanetary dust particles (IDP's) with high lateral resolution (approximately 0.2 micron), to examine possible atmospheric contamination effects, and to further explore the abilities of this technique for element analysis of small samples. The samples, previously investigated with SXRF (synchrotron X-ray fluorescence analysis), are highly enriched in Br (Br/Fe: 59 x CI, 9.2 x CI, and 116 x CI, respectively). U2015G1 is the IDP with the by far highest Zn/Fe-ratio (81 x CI) ever reported in chondritic particles.

  10. A Methodology to Measure the Cost of a Single Outpatient Visit in the Family Practice Clinic

    DTIC Science & Technology

    1986-07-01

    17,598 EDEA Other Base Support 116 " i 25,480 453 EDCA Maint Real Prop 116 Is 25,480 4,652 EDDA Minor Const 116 " H 25,480 460 EDFA Leases/Rentals 116...Engineer Liaison 116 22,319 177 EDBA Utilities 116 " " 22,319 15,414 EDEA Other Base Support 116 " " 22,319 398 EDCA Maint Real Prop 116 22,319 4,075 EDDA ...Liaison 116 4,301 34 EDBA Utilities 116 " " 4,301 2,970 EDEA Other Base Support 116 " " 4,301 76 EDCA Maint Real Prop 116 " " 4,301 785 EDDA Minor Const 116

  11. Spectral, optical and cytotoxicity studies on 2-isonicotinoyl-N-phenylhydrazine-1-carboxamide(H3L) and some of its metal complexes

    NASA Astrophysics Data System (ADS)

    Hosny, Nasser M.; Hassan, Nader Y.; Mahmoud, Heba M.; Abdel-Rhman, Mohamed H.

    2018-03-01

    The ligand 2-isonicotinoyl-N-phenylhydrazine-1-carboxamide (H3L) and its metal complexes with Co(II), Ni(II), Cu(II) and Zn(II) acetates have been synthesized. The isolated compounds have been characterized by elemental analyses, FT-IR, 1H NMR, 13C NMR, ESR, mass, electronic spectra, electrical conductivity, effective magnetic moments and thermal analyses. The free organic ligand exists in the keto form, but in the metal complexes, it coordinates in the enol form. Four coordinated species were suggested for all the isolated metal complexes. The measured optical band gap values confirmed the presence of direct electronic transition and the semi-conductivity of the compounds. The ligand and its Zn(II) complex were examined as cytotoxic agent against HCT-116 and HePG-2. The ligand showed very strong cytotoxic effect against HePG-2, but moderate cytotoxicity against HCT-116. Zn(II) complex showed weak cytotoxicity against the two cell lines.

  12. Thermophysical Properties of Matter - The TPRC Data Series. Volume 4. Specific Heat - Metallic Elements and Alloys

    DTIC Science & Technology

    1971-01-01

    alloys— sodium — sodium alloya— solder—carbon ateels—chromium steels—silicon steels—tantalum—tantalum alloys—terbium—thallium—thallium alloys—thorium...Praseodymium 45 Rhenium 46 Rhodium 47 Rubidium 48 Ruthenium 4» Samarium 50 Scandium 51 Selenium 52 Silicon 5:i Silver 54 Sodium 55 Strontium 56...Potassium ♦ Sodium 111 Sodium * Potassium 112 Tantalum ♦ Tungsten 113 Thallium + Lead, PbTl| 114 Tin ♦ Bismuth 115 Tin ♦ Indium 116 Tin+ Lead 117

  13. Exclusion Area Radiation Release during the MIT Reactor Design Basis Accident.

    DTIC Science & Technology

    1983-05-06

    Concrete Wall 116 6.2 Concrete Albedo Dose 121 6.3 Steel Door Scattering Dose 124 7.1 Total Dose Results 133 A.1 Values of N /NO for Neutron -Capture...plate fuel elements arranged in x a compact hexagonal core. This core design maximizes the neutron flux in the DO2 reflector region where numerous...sec) V = Volume of the fuel (cm 3 f Ef = Macroscopic fission cross section (cm ) = Thermal neutron flux ( neutrons /cm2 - sec) = Core-averaged value Yi

  14. Modern physics - Guest editorial

    NASA Astrophysics Data System (ADS)

    Howes, Ruth

    2000-02-01

    What an exciting time to be a physicist! You only have to attend the plenaries at an AAPT meeting to realize just how many mysteries there still are. Will string theory actually prove to be a Theory of Everything? Can we use physics to understand, predict, and perhaps eventually mitigate damaging hurricanes? Is the warming trend of the Earth due to man-made changes in the atmosphere? Have we found the nuclear "island of stability" with elements 114, 116, and 118? For that matter, why is ice slippery?

  15. Iran-Regional Country Study.

    DTIC Science & Technology

    1982-04-16

    AO-A116 226 ARMY WAR COLL CARLISLE BARRACKS PA F/6 5/4 IRAN-REGIONAL C OUNTRY STUDY.IUI UNCLASSIFIED AR8 AE I m I I..h EE f ii111112.0...ORGANIZATION NAME AND ADDRESS 10. PROGRAM ELEMENT, PROJECT. TASK AREA & WORK UNIT NUMBERS US Army War College Carlisle Barracks, PA 17013 11. CONTROLLING...PAGE(WIFhIf Date REmfot)J Bi9 US ARMY WAR COLLEGE INDIVIDUAL RESEARCH BASED ESSAY The views xpressed in this paper are those of the author and do not

  16. Application of Artificial Boundary Conditions in Sensitivity-Based Updating of Finite Element Models

    DTIC Science & Technology

    2007-06-01

    is known as the impedance matrix[ ]( )Z Ω . [ ] [ ] 1( ) ( )Z H −Ω = Ω (12) where [ ] 2( )Z K M j C ⎡ ⎤Ω = −Ω + Ω⎣ ⎦ (13) A. REDUCED ORDER...D.L. A correlation coefficient for modal vector analysis. Proceedings of 1st International Modal Analysis Conference, 1982, 110-116. Anton , H ... Rorres , C ., (2005). Elementary Linear Algebra. New York: John Wiley and Sons. Avitable, Peter (2001, January) Experimental Modal Analysis, A Simple

  17. Ligand-dependent interactions of the Ah receptor with coactivators in a mammalian two-hybrid assay

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhang Shu; Rowlands, Craig; Safe, Stephen

    2008-03-01

    2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a high affinity ligand for the aryl hydrocarbon receptor (AhR). In this study, we investigated structure-dependent differences in activation of the AhR by a series of halogenated aromatic hydrocarbons. TCDD, 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PeCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 2,3,4,7,8-pentachlorodibenzofuran (PeCDF), and 3,3',4,4',5-pentachlorobiphenyl (PCB126) induced CYP1A1-dependent activities in HEK293 human embryonic kidney, Panc1 pancreatic cancer, and Hepa1c1c7 mouse hepatoma cell lines. There was a structure-dependent difference in the efficacy of TCDF and PCB126 in HEK293 and Panc1 cells since induced CYP1A1 mRNA levels were lower than observed for the other congeners. A mammalian two-hybrid assay in cells transfected with GAL4-coactivator and AhR-VP16more » chimeras was used to investigate structure-dependent interactions of these chimeras in Panc1, HEK293, and Hepa1c1c7 cells. The reporter construct pGAL4-luc contains five tandem GAL4 response elements linked to the luciferase gene and the GAL4-coactivator chimeras express several coactivators including steroid receptor coactivator 1 (SRC-1), SRC-2 and SRC-3, the mediator coactivator TRAP220, coactivator associated arginine methyl transferase 1 (CARM-1), and peroxisome proliferator-activated receptor {gamma} coactivator 1 (PGC-1). Results of the mammalian two-hybrid studies clearly demonstrate that activation of pGAL4-luc in cells transfected with VP-AhR and GAL4-coactivator chimeras is dependent on the structure of the HAH congener, cell context, and coactivator, suggesting that the prototypical HAH congeners used in this study exhibit selective AhR modulator activity.« less

  18. Appearance of the pituitary factor Pit-1 increases chromatin remodeling at hypersensitive site III in the human GH locus.

    PubMed

    Yang, Xiaoyang; Jin, Yan; Cattini, Peter A

    2010-07-01

    Expression of pituitary and placental members of the human GH and chorionic somatomammotropin (CS) gene family is directed by an upstream remote locus control region (LCR). Pituitary-specific expression of GH requires direct binding of Pit-1 (listed as POU1F1 in the HUGO database) to sequences marked by a hypersensitive site (HS) region (HS I/II) 14.6 kb upstream of the GH-N gene (listed as GH1 in the HUGO database). We used human embryonic kidney 293 (HEK293) cells overexpressing wild-type and mutant Pit-1 proteins as a model system to gain insight into the mechanism by which Pit-1 gains access to the GH LCR. Addition of Pit-1 to these cells increased DNA accessibility at HS III, located 28 kb upstream of the human GH-N gene, in a POU homeodomain-dependent manner, as reflected by effects on histone hyperacetylation and RNA polymerase II activity. Direct binding of Pit-1 to HS III sequences is not supported. However, the potential for binding of ETS family members to this region has been demonstrated, and Pit-1 association with this ETS element in HS III sequences requires the POU homeodomain. Also, both ETS1 and ELK1 co-precipitate from human pituitary extracts using two independent sources of Pit-1 antibodies. Finally, overexpression of ELK1 or Pit-1 expression in HEK293 cells increased GH-N RNA levels. However, while ELK1 overexpression also stimulated placental CS RNA levels, the effect of Pit-1 appeared to correlate with ETS factor levels and target GH-N preferentially. These data are consistent with recruitment and an early role for Pit-1 in remodeling of the GH LCR at the constitutively open HS III through protein-protein interaction.

  19. Identification of the RNA recognition element of the RBPMS family of RNA-binding proteins and their transcriptome-wide mRNA targets

    PubMed Central

    Farazi, Thalia A.; Leonhardt, Carl S.; Mukherjee, Neelanjan; Mihailovic, Aleksandra; Li, Song; Max, Klaas E.A.; Meyer, Cindy; Yamaji, Masashi; Cekan, Pavol; Jacobs, Nicholas C.; Gerstberger, Stefanie; Bognanni, Claudia; Larsson, Erik; Ohler, Uwe; Tuschl, Thomas

    2014-01-01

    Recent studies implicated the RNA-binding protein with multiple splicing (RBPMS) family of proteins in oocyte, retinal ganglion cell, heart, and gastrointestinal smooth muscle development. These RNA-binding proteins contain a single RNA recognition motif (RRM), and their targets and molecular function have not yet been identified. We defined transcriptome-wide RNA targets using photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation (PAR-CLIP) in HEK293 cells, revealing exonic mature and intronic pre-mRNA binding sites, in agreement with the nuclear and cytoplasmic localization of the proteins. Computational and biochemical approaches defined the RNA recognition element (RRE) as a tandem CAC trinucleotide motif separated by a variable spacer region. Similar to other mRNA-binding proteins, RBPMS family of proteins relocalized to cytoplasmic stress granules under oxidative stress conditions suggestive of a support function for mRNA localization in large and/or multinucleated cells where it is preferentially expressed. PMID:24860013

  20. Direct inhibition of the DNA-binding activity of POU transcription factors Pit-1 and Brn-3 by selective binding of a phenyl-furan-benzimidazole dication.

    PubMed

    Peixoto, Paul; Liu, Yang; Depauw, Sabine; Hildebrand, Marie-Paule; Boykin, David W; Bailly, Christian; Wilson, W David; David-Cordonnier, Marie-Hélène

    2008-06-01

    The development of small molecules to control gene expression could be the spearhead of future-targeted therapeutic approaches in multiple pathologies. Among heterocyclic dications developed with this aim, a phenyl-furan-benzimidazole dication DB293 binds AT-rich sites as a monomer and 5'-ATGA sequence as a stacked dimer, both in the minor groove. Here, we used a protein/DNA array approach to evaluate the ability of DB293 to specifically inhibit transcription factors DNA-binding in a single-step, competitive mode. DB293 inhibits two POU-domain transcription factors Pit-1 and Brn-3 but not IRF-1, despite the presence of an ATGA and AT-rich sites within all three consensus sequences. EMSA, DNase I footprinting and surface-plasmon-resonance experiments determined the precise binding site, affinity and stoichiometry of DB293 interaction to the consensus targets. Binding of DB293 occurred as a cooperative dimer on the ATGA part of Brn-3 site but as two monomers on AT-rich sites of IRF-1 sequence. For Pit-1 site, ATGA or AT-rich mutated sequences identified the contribution of both sites for DB293 recognition. In conclusion, DB293 is a strong inhibitor of two POU-domain transcription factors through a cooperative binding to ATGA. These findings are the first to show that heterocyclic dications can inhibit major groove transcription factors and they open the door to the control of transcription factors activity by those compounds.

  1. Assay development of inducible human renal phosphate transporter Npt2A (SLC34A1) in Flp-In-Trex-HEK293 cells.

    PubMed

    Wu, Hongzhong; Mao, Chonghong; Duenstl, Georg; Su, Wan; Qian, Su

    2013-12-05

    Hyperphosphatemia is associated with severe decline of renal function in chronic kidney disease and elevates cardiovascular mortality. Type II sodium dependent phosphate transporter 2A (Npt2A) plays a major role in renal phosphate reabsorption and could be explored as a target for anti-hyperphosphatemia therapy. Human Npt2A transporter activity was examined upon transfection into CHO, MDCK, HEK293, Flp-In-CHO and Flp-In-HEK293 cells. Only kidney-derived cells expressed functional Npt2A. HEK293 and Flp-In-HEK293 cell lines stably transfected with hNpt2A could be selected, but these cells were inactive in phosphate transport. This suggests that high-level, constitutive Npt2A expression has deleterious effects on the cell. By using the conditional promoter in the Flp-In-Trex vector, functional expression of Npt2A was achieved by doxycycline induction in HEK293 cells. The EGFP tagged and non-tagged, inducible stable hNpt2A-HEK293 cell lines afforded development of a robust phosphate uptake assay mediated by hNpt2A, which can be used to screen hNpt2A inhibitors and inducers of hNpt2A expression. Using this assay, the small molecule LC-1 was identified as a potent inhibitor of hNpt2A, suggesting that it is feasible to develop potent specific hNpt2A inhibitors to control phosphate overloading for hyperphosphatemia therapy. © 2013 Elsevier B.V. All rights reserved.

  2. Optimization of a cAMP response element signal pathway reporter system.

    PubMed

    Shan, Qiang; Storm, Daniel R

    2010-08-15

    A sensitive cAMP response element (CRE) reporter system is essential for studying the cAMP/protein kinase A/cAMP response element binding protein signal pathway. Here we have tested a few CRE promoters and found one with high sensitivity to external stimuli. Using this optimal CRE promoter and the enhanced green fluorescent protein as the reporter, we have established a CRE reporter cell line. This cell line can be used to study the signal pathway by fluorescent microscope, fluorescence-activated cell analysis and luciferase assay. This cell line's sensitivity to forskolin, using the technique of fluorescence-activated cell sorting, was increased to approximately seven times that of its parental HEK 293 cell line, which is currently the most commonly used cell line in the field for the signal pathway study. Therefore, this newly created cell line is potentially useful for studying the signal pathway's modulators, which generally have weaker effect than its mediators. Our research has also established a general procedure for optimizing transcription-based reporter cell lines, which might be useful in performing the same task when studying many other transcription-based signal pathways. (c) 2010 Elsevier B.V. All rights reserved.

  3. Atomic weights of the elements 1999

    USGS Publications Warehouse

    Coplen, T.B.

    2001-01-01

    The biennial review of atomic-weight, Ar(E), determinations and other cognate data have resulted in changes for the standard atomic weights of the following elements: from to nitrogen 14.006 74??0.000 07 14.0067??0.0002 sulfur 32.066??0.006 32.065??0.005 chlorine 35.4527??0.0009 35.453??0.002 germanium 72.61??0.02 72.64??0.01 xenon 131.29??0.02 131.293??0.006 erbium 167.26??0.03 167.259??0.003 uranium 238.0289??0.0001 238.028 91??0.000 03 Presented are updated tables of the standard atomic weights and their uncertainties estimated by combining experimental uncertainties and terrestrial variabilities. In addition, this report again contains an updated table of relative atomic mass values and half-lives of selected radioisotopes. Changes in the evaluated isotopic abundance values from those published in 1997 are so minor that an updated list will not be published for the year 1999. Many elements have a different isotopic composition in some nonterrestrial materials. Some recent data on parent nuclides that might affect isotopic abundances or atomic-weight values are included in this report for the information of the interested scientific community. ?? 2001 American Institute of Physics.

  4. Transcriptional activation of NAD+-dependent protein deacetylase SIRT1 by nuclear receptor TLX.

    PubMed

    Iwahara, Naotoshi; Hisahara, Shin; Hayashi, Takashi; Horio, Yoshiyuki

    2009-09-04

    An orphan nuclear receptor TLX is a transcriptional repressor that promotes the proliferation and self-renewal of neural precursor cells (NPCs). SIRT1, an NAD(+)-dependent protein deacetylase, is highly expressed in the NPCs and participates in neurogenesis. Here, we found that TLX colocalized with SIRT1 and knockdown of TLX by small interfering RNAs decreased SIRT1 levels in NPCs. TLX increased the SIRT1 expression by binding to the newly identified TLX-activating element in the SIRT1 gene promoter in HEK293 cells. Thus, TLX is an inducer of SIRT1 and may contribute to neurogenesis both as a transactivator and as a repressor.

  5. Transcriptional activation of NAD{sup +}-dependent protein deacetylase SIRT1 by nuclear receptor TLX

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Iwahara, Naotoshi; Hisahara, Shin; Hayashi, Takashi

    2009-09-04

    An orphan nuclear receptor TLX is a transcriptional repressor that promotes the proliferation and self-renewal of neural precursor cells (NPCs). SIRT1, an NAD{sup +}-dependent protein deacetylase, is highly expressed in the NPCs and participates in neurogenesis. Here, we found that TLX colocalized with SIRT1 and knockdown of TLX by small interfering RNAs decreased SIRT1 levels in NPCs. TLX increased the SIRT1 expression by binding to the newly identified TLX-activating element in the SIRT1 gene promoter in HEK293 cells. Thus, TLX is an inducer of SIRT1 and may contribute to neurogenesis both as a transactivator and as a repressor.

  6. Role of Arginine 293 and Glutamine 288 in Communication between Catalytic and Allosteric Sites in Yeast Ribonucleotide Reductase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ahmad, Md. Faiz; Kaushal, Prem Singh; Wan, Qun

    2012-11-01

    Ribonucleotide reductases (RRs) catalyze the rate-limiting step of de novo deoxynucleotide (dNTP) synthesis. Eukaryotic RRs consist of two proteins, RR1 ({alpha}) that contains the catalytic site and RR2 ({beta}) that houses a diferric-tyrosyl radical essential for ribonucleoside diphosphate reduction. Biochemical analysis has been combined with isothermal titration calorimetry (ITC), X-ray crystallography and yeast genetics to elucidate the roles of two loop 2 mutations R293A and Q288A in Saccharomyces cerevisiae RR1 (ScRR1). These mutations, R293A and Q288A, cause lethality and severe S phase defects, respectively, in cells that use ScRR1 as the sole source of RR1 activity. Compared to the wild-typemore » enzyme activity, R293A and Q288A mutants show 4% and 15%, respectively, for ADP reduction, whereas they are 20% and 23%, respectively, for CDP reduction. ITC data showed that R293A ScRR1 is unable to bind ADP and binds CDP with 2-fold lower affinity compared to wild-type ScRR1. With the Q288A ScRR1 mutant, there is a 6-fold loss of affinity for ADP binding and a 2-fold loss of affinity for CDP compared to the wild type. X-ray structures of R293A ScRR1 complexed with dGTP and AMPPNP-CDP [AMPPNP, adenosine 5-({beta},{gamma}-imido)triphosphate tetralithium salt] reveal that ADP is not bound at the catalytic site, and CDP binds farther from the catalytic site compared to wild type. Our in vivo functional analyses demonstrated that R293A cannot support mitotic growth, whereas Q288A can, albeit with a severe S phase defect. Taken together, our structure, activity, ITC and in vivo data reveal that the arginine 293 and glutamine 288 residues of ScRR1 are crucial in facilitating ADP and CDP substrate selection.« less

  7. Absence of anti-hypocretin receptor 2 autoantibodies in post pandemrix narcolepsy cases.

    PubMed

    Luo, Guo; Lin, Ling; Jacob, Louis; Bonvalet, Mélodie; Ambati, Aditya; Plazzi, Giuseppe; Pizza, Fabio; Leib, Ryan; Adams, Christopher M; Partinen, Markku; Mignot, Emmanuel Jean-Marie

    2017-01-01

    A recent publication suggested molecular mimicry of a nucleoprotein (NP) sequence from A/Puerto Rico/8/1934 (PR8) strain, the backbone used in the construction of the reassortant strain X-179A that was used in Pandemrix® vaccine, and reported on anti-hypocretin (HCRT) receptor 2 (anti-HCRTR2) autoantibodies in narcolepsy, mostly in post Pandemrix® narcolepsy cases (17 of 20 sera). In this study, we re-examined this hypothesis through mass spectrometry (MS) characterization of Pandemrix®, and two other pandemic H1N1 (pH1N1)-2009 vaccines, Arepanrix® and Focetria®, and analyzed anti-HCRTR2 autoantibodies in narcolepsy patients and controls using three independent strategies. MS characterization of Pandemrix® (2 batches), Arepanrix® (4 batches) and Focetria® (1 batch) was conducted with mapping of NP 116I or 116M spectrogram. Two sets of narcolepsy cases and controls were used: 40 post Pandemrix® narcolepsy (PP-N) cases and 18 age-matched post Pandemrix® controls (PP-C), and 48 recent (≤6 months) early onset narcolepsy (EO-N) cases and 70 age-matched other controls (O-C). Anti-HCRTR2 autoantibodies were detected using three strategies: (1) Human embryonic kidney (HEK) 293T cells with transient expression of HCRTR2 were stained with human sera and then analyzed by flow cytometer; (2) In vitro translation of [35S]-radiolabelled HCRTR2 was incubated with human sera and immune complexes of autoantibody and [35S]-radiolabelled HCRTR2 were quantified using a radioligand-binding assay; (3) Optical density (OD) at 450 nm (OD450) of human serum immunoglobulin G (IgG) binding to HCRTR2 stably expressed in Chinese hamster ovary (CHO)-K1 cell line was measured using an in-cell enzyme-linked immunosorbent assay (ELISA). NP 116M mutations were predominantly present in all batches of Pandemrix®, Arepanrix® and Focetria®. The wild-type NP109-123 (ILYDKEEIRRIWRQA), a mimic to HCRTR234-45 (YDDEEFLRYLWR), was not found to bind to DQ0602. Three or four subjects were found positive for anti-HCRTR2 autoantibodies using two strategies or the third one, respectively. None of the post Pandemrix® narcolepsy cases (0 of 40 sera) was found positive with all three strategies. Anti-HCRTR2 autoantibody is not a significant biological feature of narcolepsy or of post Pandemrix® autoimmune responses.

  8. Absence of anti-hypocretin receptor 2 autoantibodies in post pandemrix narcolepsy cases

    PubMed Central

    Lin, Ling; Jacob, Louis; Bonvalet, Mélodie; Ambati, Aditya; Plazzi, Giuseppe; Pizza, Fabio; Leib, Ryan; Adams, Christopher M.; Partinen, Markku; Mignot, Emmanuel Jean-Marie

    2017-01-01

    Background A recent publication suggested molecular mimicry of a nucleoprotein (NP) sequence from A/Puerto Rico/8/1934 (PR8) strain, the backbone used in the construction of the reassortant strain X-179A that was used in Pandemrix® vaccine, and reported on anti-hypocretin (HCRT) receptor 2 (anti-HCRTR2) autoantibodies in narcolepsy, mostly in post Pandemrix® narcolepsy cases (17 of 20 sera). In this study, we re-examined this hypothesis through mass spectrometry (MS) characterization of Pandemrix®, and two other pandemic H1N1 (pH1N1)-2009 vaccines, Arepanrix® and Focetria®, and analyzed anti-HCRTR2 autoantibodies in narcolepsy patients and controls using three independent strategies. Methods MS characterization of Pandemrix® (2 batches), Arepanrix® (4 batches) and Focetria® (1 batch) was conducted with mapping of NP 116I or 116M spectrogram. Two sets of narcolepsy cases and controls were used: 40 post Pandemrix® narcolepsy (PP-N) cases and 18 age-matched post Pandemrix® controls (PP-C), and 48 recent (≤6 months) early onset narcolepsy (EO-N) cases and 70 age-matched other controls (O-C). Anti-HCRTR2 autoantibodies were detected using three strategies: (1) Human embryonic kidney (HEK) 293T cells with transient expression of HCRTR2 were stained with human sera and then analyzed by flow cytometer; (2) In vitro translation of [35S]-radiolabelled HCRTR2 was incubated with human sera and immune complexes of autoantibody and [35S]-radiolabelled HCRTR2 were quantified using a radioligand-binding assay; (3) Optical density (OD) at 450 nm (OD450) of human serum immunoglobulin G (IgG) binding to HCRTR2 stably expressed in Chinese hamster ovary (CHO)-K1 cell line was measured using an in-cell enzyme-linked immunosorbent assay (ELISA). Results NP 116M mutations were predominantly present in all batches of Pandemrix®, Arepanrix® and Focetria®. The wild-type NP109-123 (ILYDKEEIRRIWRQA), a mimic to HCRTR234-45 (YDDEEFLRYLWR), was not found to bind to DQ0602. Three or four subjects were found positive for anti-HCRTR2 autoantibodies using two strategies or the third one, respectively. None of the post Pandemrix® narcolepsy cases (0 of 40 sera) was found positive with all three strategies. Conclusion Anti-HCRTR2 autoantibody is not a significant biological feature of narcolepsy or of post Pandemrix® autoimmune responses. PMID:29220370

  9. Development of technology for modeling of a 1/8-scale dynamic model of the shuttle Solid Rocket Booster (SRB)

    NASA Technical Reports Server (NTRS)

    Levy, A.; Zalesak, J.; Bernstein, M.; Mason, P. W.

    1974-01-01

    A NASTRAN analysis of the solid rocket booster (SRB) substructure of the space shuttle 1/8-scale structural dynamics model. The NASTRAN finite element modeling capability was first used to formulate a model of a cylinder 10 in. radius by a 200 in. length to investigate the accuracy and adequacy of the proposed grid point spacing. Results were compared with a shell analysis and demonstrated relatively accurate results for NASTRAN for the lower modes, which were of primary interest. A finite element model of the full SRB was then formed using CQUAD2 plate elements containing membrane and bending stiffness and CBAR offset bar elements to represent the longerons and frames. Three layers of three-dimensional CHEXAI elements were used to model the propellant. This model, consisting of 4000 degrees of freedom (DOF) initially, was reduced to 176 DOF using Guyan reduction. The model was then submitted for complex Eigenvalue analysis. After experiencing considerable difficulty with attempts to run the complete model, it was split into two substructres. These were run separately and combined into a single 116 degree of freedom A set which was successfully run. Results are reported.

  10. Determination of elements in hospital waste with neutron activation analysis method

    NASA Astrophysics Data System (ADS)

    Dwijananti, P.; Astuti, B.; Alwiyah; Fianti

    2018-03-01

    The producer of the biggest B3 waste is hospital. The waste is from medical and laboratory activities. The purpose of this study is to determine the elements contained in the liquid waste from hospital and calculate the levels of these elements. This research was done by analysis of the neutron activation conducted at BATAN Yogyakarta. The neutron activation analysis is divided into two stages: activation of the samples using neutron sources of reactor Kartini, then chopping by using a set of tools, gamma spectrometer with HPGe detector. Qualitative and quantitative analysis were done by matching the gamma spectrum peak to the Neutron Activation Table. The sample was taken from four points of the liquid waste treatment plant (WWTP) Bhakti Wira Tamtama Semarang hospital. The results showed that the samples containing elements of Cr, Zn, Fe, Co, and Na, with the levels of each element is Cr (0.033 - 0.075) mg/L, Zn (0.090 - 1.048) mg/L, Fe (2.937-37.743) mg/L, Co (0.005-0.023) mg/L, and Na (61.088-116.330) mg/L. Comparing to the standard value, the liquid is safe to the environment.

  11. Retroviral vectors encoding ADA regulatory locus control region provide enhanced T-cell-specific transgene expression

    PubMed Central

    2009-01-01

    Background Murine retroviral vectors have been used in several hundred gene therapy clinical trials, but have fallen out of favor for a number of reasons. One issue is that gene expression from viral or internal promoters is highly variable and essentially unregulated. Moreover, with retroviral vectors, gene expression is usually silenced over time. Mammalian genes, in contrast, are characterized by highly regulated, precise levels of expression in both a temporal and a cell-specific manner. To ascertain if recapitulation of endogenous adenosine deaminase (ADA) expression can be achieved in a vector construct we created a new series of Moloney murine leukemia virus (MuLV) based retroviral vector that carry human regulatory elements including combinations of the ADA promoter, the ADA locus control region (LCR), ADA introns and human polyadenylation sequences in a self-inactivating vector backbone. Methods A MuLV-based retroviral vector with a self-inactivating (SIN) backbone, the phosphoglycerate kinase promoter (PGK) and the enhanced green fluorescent protein (eGFP), as a reporter gene, was generated. Subsequent vectors were constructed from this basic vector by deletion or addition of certain elements. The added elements that were assessed are the human ADA promoter, human ADA locus control region (LCR), introns 7, 8, and 11 from the human ADA gene, and human growth hormone polyadenylation signal. Retroviral vector particles were produced by transient three-plasmid transfection of 293T cells. Retroviral vectors encoding eGFP were titered by transducing 293A cells, and then the proportion of GFP-positive cells was determined using fluorescence-activated cell sorting (FACS). Non T-cell and T-cell lines were transduced at a multiplicity of infection (MOI) of 0.1 and the yield of eGFP transgene expression was evaluated by FACS analysis using mean fluorescent intensity (MFI) detection. Results Vectors that contained the ADA LCR were preferentially expressed in T-cell lines. Further improvements in T-cell specific gene expression were observed with the incorporation of additional cis-regulatory elements, such as a human polyadenylation signal and intron 7 from the human ADA gene. Conclusion These studies suggest that the combination of an authentically regulated ADA gene in a murine retroviral vector, together with additional locus-specific regulatory refinements, will yield a vector with a safer profile and greater efficacy in terms of high-level, therapeutic, regulated gene expression for the treatment of ADA-deficient severe combined immunodeficiency. PMID:20042112

  12. Reduction of adenovirus E1A mRNA by RNAi results in enhanced recombinant protein expression in transiently transfected HEK293 cells.

    PubMed

    Hacker, David L; Bertschinger, Martin; Baldi, Lucia; Wurm, Florian M

    2004-10-27

    Human embryonic kidney 293 (HEK293) cells, a widely used host for large-scale transient expression of recombinant proteins, are transformed with the adenovirus E1A and E1B genes. Because the E1A proteins function as transcriptional activators or repressors, they may have a positive or negative effect on transient transgene expression in this cell line. Suspension cultures of HEK293 EBNA (HEK293E) cells were co-transfected with a reporter plasmid expressing the GFP gene and a plasmid expressing a short hairpin RNA (shRNA) targeting the E1A mRNAs for degradation by RNA interference (RNAi). The presence of the shRNA in HEK293E cells reduced the steady state level of E1A mRNA up to 75% and increased transient GFP expression from either the elongation factor-1alpha (EF-1alpha) promoter or the human cytomegalovirus (HCMV) immediate early promoter up to twofold. E1A mRNA depletion also resulted in a twofold increase in transient expression of a recombinant IgG in both small- and large-scale suspension cultures when the IgG light and heavy chain genes were controlled by the EF-1alpha promoter. Finally, transient IgG expression was enhanced 2.5-fold when the anti-E1A shRNA was expressed from the same vector as the IgG light chain gene. These results demonstrated that E1A has a negative effect on transient gene expression in HEK293E cells, and they established that RNAi can be used to enhance recombinant protein expression in mammalian cells.

  13. Investigation of free fatty acid associated recombinant membrane receptor protein expression in HEK293 cells using Raman spectroscopy, calcium imaging, and atomic force microscopy.

    PubMed

    Lin, Juqiang; Xu, Han; Wu, Yangzhe; Tang, Mingjie; McEwen, Gerald D; Liu, Pin; Hansen, Dane R; Gilbertson, Timothy A; Zhou, Anhong

    2013-02-05

    G-protein-coupled receptor 120 (GPR120) is a previously orphaned G-protein-coupled receptor that apparently functions as a sensor for dietary fat in the gustatory and digestive systems. In this study, a cDNA sequence encoding a doxycycline (Dox)-inducible mature peptide of GPR120 was inserted into an expression vector and transfected in HEK293 cells. We measured Raman spectra of single HEK293 cells as well as GPR120-expressing HEK293-GPR120 cells at a 48 h period following the additions of Dox at several concentrations. We found that the spectral intensity of HEK293-GPR120 cells is dependent upon the dose of Dox, which correlates with the accumulation of GPR120 protein in the cells. However, the amount of the fatty acid activated changes in intracellular calcium (Ca(2+)) as measured by ratiometric calcium imaging was not correlated with Dox concentration. Principal components analysis (PCA) of Raman spectra reveals that the spectra from different treatments of HEK293-GPR120 cells form distinct, completely separated clusters with the receiver operating characteristic (ROC) area of 1, while those spectra for the HEK293 cells form small overlap clusters with the ROC area of 0.836. It was also found that expression of GPR120 altered the physiochemical and biomechanical properties of the parental cell membrane surface, which was quantitated by atomic force microscopy (AFM). These findings demonstrate that the combination of Raman spectroscopy, calcium imaging, and AFM may provide new tools in noninvasive and quantitative monitoring of membrane receptor expression induced alterations in the biophysical and signaling properties of single living cells.

  14. Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells.

    PubMed

    Böhm, Ernst; Seyfried, Birgit K; Dockal, Michael; Graninger, Michael; Hasslacher, Meinhard; Neurath, Marianne; Konetschny, Christian; Matthiessen, Peter; Mitterer, Artur; Scheiflinger, Friedrich

    2015-09-18

    BACKGROUND & Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. RESULTS & The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII.

  15. Utility of human embryonic kidney cell line HEK-293 for rapid isolation of fixed and street rabies viruses: comparison with Neuro-2a and BHK-21 cell lines.

    PubMed

    Madhusudana, Shampur Narayan; Sundaramoorthy, Subha; Ullas, Padinjaremattatthil Thankappan

    2010-12-01

    A confirmatory rabies diagnosis can be achieved by rapid virus isolation in cell culture using brain tissue from the suspect animal. Several cell lines have been used for this purpose and the murine neuroblastoma cell line Neuro-2a has been found to be the most sensitive. The human embryonic kidney cell line HEK-293 is known to express several neuronal proteins and is believed to be of neuronal origin. We hypothesized that this cell line could be susceptible to rabies virus, which is highly neurotropic. First we tested the sensitivity of HEK-293 cells to the laboratory strain, challenge virus standard (CVS). We then tested 120 brain samples from different animals and humans suspected to have died of rabies by fluorescent antibody test (FAT). Both FAT-positive and FAT-negative brains were tested for virus isolation using Neuro-2a, BHK-21, and HEK-293 cell lines and also by mouse inoculation. There was 100% correlation between FAT, virus isolation in Neuro-2a and HEK-293 cells, and mouse inoculation. However, the rate of virus isolation in the BHK-21 cell line was only 28% when compared to the other cell lines. The sensitivity of HEK-293 to CVS strain of virus was similar to that of Neuro-2a. We conclude that the HEK-293 cell line is as sensitive as the Neuro-2a cell line for the rapid isolation of rabies virus and may serve as an alternative cell line for rabies diagnosis and future research. Copyright © 2010 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  16. [Construction of Rev-erbβ gene knockout HEK293 cell line with CRISPR/Cas9 system].

    PubMed

    Chen, Fang; Zhang, Weifeng; Zhao, Junli; Yang, Peiyan; Ma, Rui; Xia, Haibin

    2016-11-01

    Objective To prepare Rev-erbβ knockout HEK293 cells using clustered regularly interspaced short palindromic repeats/Cas 9 nuclease (CRISPR/Cas9) gene editing technology. Methods The knock-in or knockout of Rev-erbβ gene could be realized by single-guide RNA (sgRNA)-mediated Cas9 cutting of target DNA, and followed by DNA homologous recombination or non-homologous end joining-mediated DNA repair. Firstly, four sgRNAs were designed for Rev-erbβ gene. The sgRNA1 and sgRNA2 with the higher activity were respectively used to construct pCMV-hCas9-U6-Rev-erbβ sgRNA1 and pCMV-hCas9-U6-Rev-erbβ sgRNA2. Then, pCMV-hCas9-U6-Rev-erbβ sgRNA1, pCMV-hCas9-U6-Rev-erbβ sgRNA2 and pAd5-E1/hRev-erbβ donor plasmid vectors were co-transfected into HEK293 cells. Through drug screening, cloning and sequencing, the Rev-erbβ gene-knockout HEK293 (Rev-erbβ -/- ) cell lines were obtained with one chain integrated with exogenous gene fragment and the other chain for deletion mutants. Finally, the HEK293 (Rev-erbβ -/- ) cell lines (C3-6) was detected with real-time quantitative PCR and Western blotting. Results Expression of Rev-erbβ mRNA and protein was undetectable in HEK293 Rev-erbβ -/- cell line. Conclusion Using CRISPR/Cas9 technology, the HEK293 Rev-erbβ -/- cell line has been successfully constructed, which would provide an effective tool for the study on the function of Rev-erbβ.

  17. Buckling Testing and Analysis of Honeycomb Sandwich Panel Arc Segments of a Full-Scale Fairing Barrel. Part 3; 8-ply Out-of-Autoclave Facesheets

    NASA Technical Reports Server (NTRS)

    Pineda, Evan J.; Myers, David E.; Kosareo, Daniel N.; Kellas, Sotiris

    2014-01-01

    Four honeycomb sandwich panels, representing 1/16th arc segments of a 10 m diameter barrel section of the heavy lift launch vehicle, were manufactured under the NASA Composites for Exploration program and the NASA Constellation Ares V program. Two configurations were chosen for the panels: 6-ply facesheets with 1.125 in. honeycomb core and 8-ply facesheets with 1.000 in. honeycomb core. Additionally, two separate carbon fiber/epoxy material systems were chosen for the facesheets: inautoclave IM7/977-3 and out-of-autoclave T40-800B/5320-1. Smaller 3- by 5-ft panels were cut from the 1/16th barrel sections. These panels were tested under compressive loading at the NASA Langley Research Center. Furthermore, linear eigenvalue and geometrically nonlinear finite element analyses were performed to predict the compressive response of the 3- by 5-ft panels. This manuscript summarizes the experimental and analytical modeling efforts pertaining to the panel composed of 8-ply, T40-800B/5320-1 facesheets (referred to as Panel C). To improve the robustness of the geometrically nonlinear finite element model, measured surface imperfections were included in the geometry of the model. Both the linear and nonlinear, two-dimensional (2-D) and three-dimensional (3-D), models yield good qualitative and quantitative predictions. Additionally, it was predicted correctly that the panel would fail in buckling prior to failing in strength.

  18. The role of Ia molecules in the activation of T lymphocytes. I. The activation of an IL 1-dependent IL 2-producing T cell hybridoma by Con A requires an interaction, which is not H-2-restricted, with an Ia-bearing accessory cell.

    PubMed

    Rock, K L

    1982-10-01

    A model of accessory cell-dependent lectin-mediated T cell activation was investigated by utilizing a mitogen-inducible T cell hybridoma. A continuous MHC-restricted antigen-specific T cell line was fused with the azaguanine-resistant AKR thymoma BW5147. A hybrid, RF1.16B, was identified that is minimally inducible by Con A stimulation alone but is stimulated by Con A in the presence of T cell-depleted accessory cells to produce interleukin 2. The accessory cell function can be replaced by the monokine interleukin 1. Thus the lectin is a sufficient trigger for the hybrid in the absence of MHC restriction elements. The accessory cell function from splenocytes is provided by a non-B, non-T, predominantly Ia-bearing radioresistant cell. The interaction between the RF1.16B hybrid and the accessory cell population is not H-2-restricted. Control experiments, including the use of a cloned source of accessory cells, ruled out contaminating T cells or direct lectin effects as an explanation for the lack of H-2 restriction. The finding that an Ia-bearing cell is required for activation in an MHC-nonrestricted manner is discussed, and a hypothesis is raised that Ia antigens may play a role in addition to that of being a restriction element.

  19. Implementation and Evaluation of a Recurring Interdisciplinary Community Health Fair in a Remote U.S.-Mexico Border Community.

    PubMed

    Lee, July; McKennett, Marianne; Rodriguez, Xavier; Smith, Sunny

    2018-03-06

    The purpose of this project was to design, implement, and assess a recurring interdisciplinary community health fair in an underserved border town. University of California San Diego (UCSD) medical and pharmacy students, under faculty supervision, worked alongside community partners in Calexico, California to implement a health fair two miles from the U.S.-Mexico border. Demographic and screening data were described from 293 participants from 2014 to 2016. Over 90% (269/293) listed Mexico as their country of birth, 82.9% (243/293) were monolingual Spanish speakers, 75.4% (221/293) had an annual household income of ≤ $20,000, and 58.7% (172/293) described their health as fair or poor. Screening revealed 91.1% (265/291) were overweight or obese, 37.8% (109/288) had hypertension, 9.3% (27/289) had elevated blood sugar, and 11.4% (33/289) had elevated total cholesterol levels. This model could be replicated in other training settings to increase exposure to border health issues and connect patients to local health services.

  20. Human Embryonic Kidney 293 Cells: A Vehicle for Biopharmaceutical Manufacturing, Structural Biology, and Electrophysiology.

    PubMed

    Hu, Jianwen; Han, Jizhong; Li, Haoran; Zhang, Xian; Liu, Lan Lan; Chen, Fei; Zeng, Bin

    2018-01-01

    Mammalian cells, e.g., CHO, BHK, HEK293, HT-1080, and NS0 cells, represent important manufacturing platforms in bioengineering. They are widely used for the production of recombinant therapeutic proteins, vaccines, anticancer agents, and other clinically relevant drugs. HEK293 (human embryonic kidney 293) cells and their derived cell lines provide an attractive heterologous system for the development of recombinant proteins or adenovirus productions, not least due to their human-like posttranslational modification of protein molecules to provide the desired biological activity. Secondly, they also exhibit high transfection efficiency yielding high-quality recombinant proteins. They are easy to maintain and express with high fidelity membrane proteins, such as ion channels and transporters, and thus are attractive for structural biology and electrophysiology studies. In this article, we review the literature on HEK293 cells regarding their origins but also stress their advancements into the different cell lines engineered and discuss some significant aspects which make them versatile systems for biopharmaceutical manufacturing, drug screening, structural biology research, and electrophysiology applications. © 2018 S. Karger AG, Basel.

  1. Tn5401, a new class II transposable element from Bacillus thuringiensis.

    PubMed Central

    Baum, J A

    1994-01-01

    A new class II (Tn3-like) transposable element, designated Tn5401, was recovered from a sporulation-deficient variant of Bacillus thuringiensis subsp. morrisoni EG2158 following its insertion into a recombinant plasmid. Sequence analysis of the insert revealed a 4,837-bp transposon with two large open reading frames, in the same orientation, encoding proteins of 36 kDa (306 residues) and 116 kDa (1,005 residues) and 53-bp terminal inverted repeats. The deduced amino acid sequence for the 36-kDa protein shows 24% sequence identity with the TnpI recombinase of the B. thuringiensis transposon Tn4430, a member of the phage integrase family of site-specific recombinases. The deduced amino acid sequence for the 116-kDa protein shows 42% sequence identity with the transposase of Tn3 but only 28% identity with the TnpA transposase of Tn4430. Two small open reading frames of unknown function, designated orf1 (85 residues) and orf2 (74 residues), were also identified. Southern blot analysis indicated that Tn5401, in contrast to Tn4430, is not commonly found among different subspecies of B. thuringiensis and is not typically associated with known insecticidal crystal protein genes. Transposition was studied with B. thuringiensis by using plasmid pEG922, a temperature-sensitive shuttle vector containing Tn5401. Tn5401 transposed to both chromosomal and plasmid target sites but displayed an apparent preference for plasmid sites. Transposition was replicative and resulted in the generation of a 5-bp duplication at the target site. Transcriptional start sites within Tn5401 were mapped by primer extension analysis. Two promoters, designated PL and PR, direct the transcription of orf1-orf2 and tnpI-tnpA, respectively, and are negatively regulated by TnpI. Sequence comparison of the promoter regions of Tn5401 and Tn4430 suggests that the conserved sequence element ATGTCCRCTAAY mediates TnpI binding and cointegrate resolution. The same element is contained within the 53-bp terminal inverted repeats, thus accounting for their unusual lengths and suggesting an additional role for TnpI in regulating Tn5401 transposition. Images PMID:7514590

  2. Effects of site-directed mutagenesis of Asn116 in the β-hairpin of the N-terminal domain of thermolysin on its activity and stability.

    PubMed

    Menach, Evans; Yasukawa, Kiyoshi; Inouye, Kuniyo

    2012-09-01

    In the N-terminal domain of thermolysin, two anti-parallel β-strands, Asn112-Ala113-Phe114-Trp115 and Ser118-Gln119-Met120-Val121-Tyr122 are connected by an Asn116-Gly117 turn to form a β-hairpin structure. In this study, we examined the role of Asn116 in the activity and stability of thermolysin by site-directed mutagenesis. Of the 19 Asn116 variants, four (N116A, N116D, N116T and N116Q) were produced in Escherichia coli, by co-expressing the mature and pro domains separately, while the other 15 were not. In the hydrolysis of N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide (FAGLA) at 25°C, the intrinsic k(cat)/K(m) value of N116D was 320% of that of the wild-type thermolysin (WT), and in the hydrolysis of N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester (ZDFM) at pH 7.5 at 25°C, the k(cat)/K(m) value of N116D was 140% of that of WT, indicating that N116D exhibited higher activity than WT. N116Q exhibited similar activity as WT, and N116A and N116T exhibited reduced activities. The first-order rate constants, k(obs), of the thermal inactivation at 80°C were in the order N116A, N116D, N116T > N116Q > WT at all CaCl(2) concentrations examined (1-100 mM), indicating that all variants exhibited reduced stabilities. These results suggest that Asn116 plays an important role in the activity and stability of thermolysin presumably by stabilizing this β-hairpin structure.

  3. Synthesis, characterization, and anticancer activity of a series of ketone-N(4)-substituted thiosemicarbazones and their ruthenium(II) arene complexes.

    PubMed

    Su, Wei; Qian, Quanquan; Li, Peiyuan; Lei, Xiaolin; Xiao, Qi; Huang, Shan; Huang, Chusheng; Cui, Jianguo

    2013-11-04

    A series of ketone-N(4)-substituted thiosemicarbazone (TSC) compounds (L1-L9) and their corresponding [(η(6)-p-cymene)Ru(II)(TSC)Cl](+/0) complexes (1-9) were synthesized and characterized by NMR, IR, elemental analysis, and HR-ESI-mass spectrometry. The molecular structures of L4, L9, 1-6, and 9 were determined by single-crystal X-ray diffraction analysis. The compounds were further evaluated for their in vitro antiproliferative activities against the SGC-7901 human gastric cancer, BEL-7404 human liver cancer, and HEK-293T noncancerous cell lines. Furthermore, the interactions of the compounds with DNA were followed by electrophoretic mobility spectrometry studies.

  4. Vulcanism, mercury-sensitized photo-reactions and abiogenetic synthesis - A theoretical treatment

    NASA Technical Reports Server (NTRS)

    Siegel, S. M.; Siegel, B. Z.

    1976-01-01

    Attention is called to the photodynamic and thermodynamic properties of Periodic Group IIb elements, most notably Hg, as they relate to ultra-violet sensitization in organic chemical reactions. The energy levels of 6 1P1 and 6 3P1 resonance states and the high vapor pressure (greater than 0.001 mm) of the metal at temperatures as low as 293 K bring Hg with the range of bond dissociation energies in most organic molecules and many inorganics. These capabilities considered together with recent evidence for Hg emission as a regular part of volcanic and geothermal processes provide the basis for a proposal that Hg-sensitized ultraviolet photo-reactions may have played a significant part in abiogenetic organic synthesis on the primative earth.

  5. Evaluation of the Utility of Static and Adaptive Mesh Refinement for Idealized Tropical Cyclone Problems in a Spectral Element Shallow Water Model

    DTIC Science & Technology

    2015-04-09

    where u is the zonal momentum per unit mass, v is the meridional momentum per unit mass, h is the fluid depth, and f is the Coriolis parameter. An...from each cyclone advects the other116 creating a net cyclonic motion (the Fujiwhara effect ; Fujiwhara 1921) (case 2 idealization).117 In Fig. 2c, the...the interaction of the two136 vortices cause a net cyclonic motion (the Fujiwhara effect ).137 The initial condition for the binary vortex interaction

  6. Thiosulfate as an Antidote to Mustard Poisoning, A Review of the Literature.

    DTIC Science & Technology

    1982-09-01

    FRANCISCO CA UNCLASSIFIED K D MCKINLEY ET AL. SEP 82 LRIR-127 F/G 61209 N MENOMONEE NONIEhhhhhhhhhEND 1168 UL 1.0. * 1.25 I 1,1.6 MICROCOPY RESOLUTION TEST...ORGANIZATION NAME AND ADDRESS 10. PRORA ELEMENT. PROJECT, TASK AREA &J’WORC UNIT N UMSIERS Division of Research Support, Letterman ArmyPojN/3124A7 Institute...block 2,If different fram Report) I& SUPPLI111ENTARY NOTES I9. KEY WORDS (Centinue n reverse side If natessy and identlity5 b ock number) Sodium

  7. Thermal Conductivity of Metals and Alloys at Low Temperatures. A Review of the Literature

    DTIC Science & Technology

    1954-09-01

    between OnF and Ag , Ni. anda .. er.ta.. ducovruml s f (96) .. J m Sa . Oye-rveehg odciiy.. W.1 Powe28). sample .. &...... - .3a. IM. C.. Dp..... .Je...R. KikuchiBERYLIUM (932). Cominrl See figure 2, under "Metallic Elements".. E.J. Lewi 6.0 Ag ............. k= 1.16at27WC...9 Al 2 Zn, 0.1 .71 1.32 ........................ Aged . Mo. U ~5.Cu, 0.5 Si, 0.8 Fe, 0.5 Mn, 1.23 ......................... Quenched. MIA.B

  8. The Effect of Aviation Fuels Containing Low Amounts of Static Dissipater Additive on Electrostatic Charge Generation

    DTIC Science & Technology

    1994-05-01

    60 9.44 22.67 105 53.3 3.4 11.5 70 963 22.6? 12: so 3.14 11 S so 9.73 22.57 135 Soo 3 4 116 s0 9.73 22.6? ISO 53.3 3.4 11.6 t00 9.75 22.67 165...50.0 3.4 11.6 I10 9.73 22.57 ISO 5O0. 3.4 11.6 120 9.73 22.67 115 50.0 3.4 11.6 130 9.76 22.67 210 50.0 3.4 11.6 140 9.73 22.67 225 S3.3 3.4 11.6I... ISO 9.75 22.67 240 50.0 3.4 11.6 160 9.67 22.67 255 50.0 3.4 11.6 170 I.53 22.67 270 50.* 3.4 11.6 Igo 9.46 22.67 265 5O0. 3.4 11.6U Ito 9.03 22.67

  9. Efficient synthesis and evaluation of bis-pyridinium/bis-quinolinium metallosalophens as antibiotic and antitumor candidates

    NASA Astrophysics Data System (ADS)

    Elshaarawy, Reda F. M.; Eldeen, Ibrahim M.; Hassan, Eman M.

    2017-01-01

    Inspired with the pharmacological diversity of salophens and in our endeavor to explore a new strategy which may conflict the invasion of drug resistance, we report herein efficient synthetic routes for the synthesis of new RO-salophen(Cl), pyridinium/quinolinium-based salophens (3a-e) and metallosalophens (4a-j). These new architectures have been structurally characterized by elemental and spectral analysis as well pharmacologically evaluated for their in vitro antimicrobial, against a common panel of pathogenic bacterial and fungal strains, and anticancer activities against human colon carcinoma (HCT-116) cell lines. Antimicrobial assay results revealed that all tested compounds exhibited moderate to superb broad-spectrum efficacy in comparison to the standard antibiotic with a preferential ability to perform as a fungicides than to act as bactericides. Noteworthy, VO(II)-salophens are more effective in reduction HCT-116 cell viability than Cu(II)-salophens. For example, VO(II)-salophen3 (4f) (IC50 = 2.13 μg/mL) was ca. 10-fold more efficient than Cu(II)-salophen3 (4e) (IC50 = 20.30 μg/mL).

  10. Atomic Weights of the Elements 1999

    NASA Astrophysics Data System (ADS)

    Coplen, T. B.

    2001-05-01

    The biennial review of atomic-weight, Ar(E), determinations and other cognate data have resulted in changes for the standard atomic weights of the following elements: from to nitrogen 14.006 74±0.000 07¯r 14.0067±0.0002¯ sulfur 32.066±0.006 32.065±0.005 chlorine 35.4527±0.0009 35.453±0.002 germanium 72.61±0.02 72.64±0.01 xenon 131.29±0.02 131.293±0.006 erbium 167.26±0.03 167.259±0.003 uranium 238.0289±0.0001 238.028 91±0.000 03 Presented are updated tables of the standard atomic weights and their uncertainties estimated by combining experimental uncertainties and terrestrial variabilities. In addition, this report again contains an updated table of relative atomic mass values and half-lives of selected radioisotopes. Changes in the evaluated isotopic abundance values from those published in 1997 are so minor that an updated list will not be published for the year 1999. Many elements have a different isotopic composition in some nonterrestrial materials. Some recent data on parent nuclides that might affect isotopic abundances or atomic-weight values are included in this report for the information of the interested scientific community.

  11. Trace element fluxes in sediments of an environmentally impacted river from a coastal zone of Brazil.

    PubMed

    da Silva, Yuri Jacques Agra Bezerra; Cantalice, José Ramon Barros; Singh, Vijay P; do Nascimento, Clístenes Williams Araújo; Piscoya, Victor Casimiro; Guerra, Sérgio M S

    2015-10-01

    Data regarding trace element concentrations and fluxes in suspended sediments and bedload are scarce. To fill this gap and meet the international need to include polluted rivers in future world estimation of trace element fluxes, this study aimed to determine the trace element fluxes in suspended sediment and bedload of an environmentally impacted river in Brazil. Water, suspended sediment, and bedload from both the upstream and the downstream cross sections were collected. To collect both the suspended sediment and water samples, we used the US DH-48. Bedload measurements were carried out using the US BLH 84 sampler. Concentrations of Cd, Cr, Cu, Fe, Mn, Ni, Pb, and Zn were determined by inductively coupled plasma (ICP-OES). As and Hg were determined by an atomic absorption spectrophotometer (AA-FIAS). The suspended sediments contributed more than 99 % of the trace element flux. By far Pb and to a less extent Zn at the downstream site represents major concerns. The yields of Pb and Zn in suspended sediments were 4.20 and 2.93 kg km(2) year(-1), respectively. These yields were higher than the values reported for Pb and Zn for Tuul River (highly impacted by mining activities), 1.60 and 1.30 kg km(2) year(-1), respectively, as well as the Pb yield (suspended + dissolved) to the sea of some Mediterranean rivers equal to 3.4 kg km(2) year(-1). Therefore, the highest flux and yield of Pb and Zn in Ipojuca River highlighted the importance to include medium and small rivers-often overlooked in global and regional studies-in the future estimation of world trace element fluxes in order to protect estuaries and coastal zones.

  12. The Proteins from Sika deer antler as potential modulators on cisplatin-induced cytotoxicity in human embryonic kidney 293 cells.

    PubMed

    Yang, Huihai; Li, Wei; Wang, Lulu; He, Xiaofeng; Sun, Hang; Zhang, Jing

    2017-07-31

    Our study aimed to investigate the protective role of SDAPR on cisplatin-induced cytotoxicity and its' possible mechanism in HEK293 cells. Cell viability was measured by MTT assay. Oxidative stress (SOD, GSH, LDH and MDA), inflammatory factors (TNF-α and IL-6) and apoptosis-related proteins (caspase-3, Bax, Bcl-2) expression were measured. The apoptotic cells were observed by TUNEL staining. Our study results indicated that non-cytotoxic levels of SDAPR significantly increased viability rate (LD 50 value of cisplatin is 20 μM), which improved antioxidant defence, attenuated apoptosis by decreasing expression levels of cleaved-caspase-3 and Bax, increasing Bcl-2 expression and inhibiting apoptotic positive cells in HEK 293 cells. In addition, SDAPR treatment markedly inhibited the levels of TNF-α and IL-6. In conclusion, Sika deer antler protein, a potential modulator, could alleviate cisplatin-induced cytotoxicity in HEK 293 cells.

  13. Comparison of two eukaryotic systems for the expression of VP6 protein of rotavirus specie A: transient gene expression in HEK293-T cells and insect cell-baculovirus system.

    PubMed

    da Silva Junior, Haroldo Cid; da Silva E Mouta Junior, Sérgio; de Mendonça, Marcos César Lima; de Souza Pereira, Mirian Claudia; da Rocha Nogueira, Alanderson; de Azevedo, Maria Luiza Borges; Leite, José Paulo Gagliardi; de Moraes, Márcia Terezinha Baroni

    2012-09-01

    The VP6 protein of rotavirus A (RVA) is a target antigen used for diagnostic assays and also for the development of new RVA vaccines. We have compared the expression of VP6 protein in human embryonic kidney (HEK293-T) cells with results obtained using a well-established insect cell-baculovirus system. The recombinant VP6 (rVP6) expressed in HEK293-T cells did not present degradation and also retained the ability to form trimers. In the insect cell-baculovirus system, rVP6 was expressed at higher levels and with protein degradation as well as partial loss of ability to form trimers was observed. Therefore, HEK293-T cells represent a less laborious alternative system than insect cells for expression of rVP6 from human RVA.

  14. Seaweeds along KwaZulu-Natal coast of South Africa-3: elemental uptake by Ulva lactuca (Sea lettuce).

    PubMed

    Misheer, Natasha; Kindness, A; Jonnalagadda, S B

    2006-01-01

    The elemental uptake by Ulva lactuca (Sea lettuce), a marine macro-algae (chlorophyta, green alga) grown richly along KwaZulu-Natal coastline. The total elemental concentrations of seven important elements, namely manganese, iron, arsenic, boron, titanium, zinc and mercury, selected based on their abundance in U. lactuca were investigated for one year cycle (June 2002 to May 2003). The four selected sampling sites, Zinkwasi, Ballito, Treasure Beach and Park Ryrie are spread over 150 km wide along the KwaZulu-Natal coastline from North to South. The Ulva lectuca possess good manganese and arsenic accumulating ability and an excellent bio-indicator for most of the metals studied. A typical U. lectuca sample at Zinkwasi (in winter) recorded Mn (25.3 +/- 1.16 ppm), Fe (21.0 +/- 0.85 ppm), As (6.2 +/- 0.30 ppm), B (935 +/- 14 ppb), Ti (863 +/- 34 ppb), Zn (421 +/- 21 ppb), and Hg (61.3 +/- 1.2 ppb). The general trend found at all sites was high elemental concentrations in winter and a decrease in concentrations from winter to spring and summer. Iron uptake was lowest in summer and autumn at all sites. Ulva lactuca recorded highest mercury levels (>400 ppb) during the spring season at the Treasure Beach site near Durban.

  15. Oxygen concentrators performance with nitrous oxide at 50:50 volume.

    PubMed

    Moll, Jorge Ronaldo; Vieira, Joaquim Edson; Gozzani, Judymara Lauzi; Mathias, Lígia Andrade Silva Telles

    2014-01-01

    Few investigations have addressed the safety of oxygen from concentrators for use in anesthesia in association with nitrous oxide. This study evaluated the percent of oxygen from a concentrator in association with nitrous oxide in a semi-closed rebreathing circuit. Adult patients undergoing low risk surgery were randomly allocated into two groups, receiving a fresh gas flow of oxygen from concentrators (O293) or of oxygen from concentrators and nitrous oxide (O293N2O). The fraction of inspired oxygen and the percentage of oxygen from fresh gas flow were measured every 10 min. The ratio of FiO2/oxygen concentration delivered was compared at various time intervals and between the groups. Thirty patients were studied in each group. There was no difference in oxygen from concentrators over time for both groups, but there was a significant improvement in the FiO2 (p<0.001) for O293 group while a significant decline (p<0.001) for O293N2O. The FiO2/oxygen ratio varied in both groups, reaching a plateau in the O293 group. Pulse oximetry did not fall below 98.5% in either group. The FiO2 in the mixture of O293 and nitrous oxide fell during the observation period although oxygen saturation was higher than 98.5% throughout the study. Concentrators can be considered a stable source of oxygen for use during short anesthetic procedures, either pure or in association with nitrous oxide at 50:50 volume. Copyright © 2013 Sociedade Brasileira de Anestesiologia. Published by Elsevier Editora Ltda. All rights reserved.

  16. Anisotropic Thermal Expansion of Zirconium Diboride: An Energy-Dispersive X-Ray Diffraction Study

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Paxton, William A.; Özdemir, Tevfik E.; Şavklıyıldız, İlyas

    Zirconiumore » m diboride (ZrB 2) is an attractive material due to its thermal and electrical properties. In recent years, ZrB 2has been investigated as a superior replacement for sapphire when used as a substrate for gallium nitride devices. Like sapphire, ZrB 2has an anisotropic hexagonal structure which defines its directionally dependent properties. However, the anisotropic behavior of ZrB 2is not well understood. In this paper, we use energy-dispersive synchrotron X-ray diffraction to measure the thermal expansion of polycrystalline ZrB 2powder from 300 to 1150 K. Nine Bragg reflections are fit using Pseudo-Voigt peak profiles and used to compute the a and c lattice parameters using a nonlinear least-squares approximation. The temperature-dependent instantaneous thermal expansion coefficients are determined for each a -axis and c -axis direction and are described by the following equations: α a = ( 4.1507 × 10 - 6 + 5.1086 × 10 - 9 ( T - 293.15 ) )/( 1 + 4.1507 × 10 - 6 ( T - 293.15 ) + 2.5543 × 10 - 9 ( T - 293.15 ) 2 ) and α c = ( 4.5374 × 10 - 6 + 4.3004 × 10 - 9 ( T - 293.15 ) )/( 1 + 4.5374 × 10 - 6 ( T - 293.15 ) + 2.1502 × 10 - 9 ( T - 293.15 ) 2 ). Our results are within range of previously reported values but describe the temperature anisotropy in more detail. We show that anisotropic expansion coefficients converge to the same value at about 780 K and diverge at higher temperatures. Results are compared with other reported values.« less

  17. Determination of trace elements in epiphytic lichens from Bandar Baru Bangi, Selangor using INAA method

    NASA Astrophysics Data System (ADS)

    Khairudin, Nurshafiq Ezam; Siong, Khoo Kok; Siong, Wee Boon

    2014-02-01

    Lichens have been used as effective biomonitors of atmospheric pollutants as they can take up nutrients and pollutants directly from the atmosphere. In this study, trace element contents in epiphytic lichens were determined using INAA method. Samples were collected from 7 sampling locations around Bandar Baru Bangi, Selangor. The elements detected were As (1.73+0.85 mg/kg), Ce (3.65+1.91 mg/kg), Co (0.29+0.12 mg/kg), Cr (5.92+3.54 mg/kg), Cs (0.92+0.25 mg/kg), Eu (0.03+0.02 mg/kg), Fe (1280+760 mg/kg), Hf (0.37+0.18 mg/kg), La (1.52+0.89 mg/kg), Rb (27.7+4.8 mg/kg), Sc (0.33+0.19 mg/kg), Sm (0.28+0.16 mg/kg), Th (1.21+0.62 mg/kg) and Zn (116+27 mg/kg). Comparisons were then made between the elemental concentrations obtained and the baseline data from literature. Results showed that most of the elements were within the concentration range of the baseline data. Enrichment factors (EF) of the trace element in lichens showed that most of the elements were within the range of the baseline data except for As which was found to be slightly enriched (EF: 13.2 - 28.5). Regression analysis indicated significant correlation (p<0.05) with Sc for most of the elements which signifies crustal input except for Cs and Rb. The poor correlations of Cs and Rb with Sc may be due to the mobility of these elements. In summary, trace element data obtained using INAA were very useful and demonstrated that lichens were suitable biomonitors for identifying potential trace element pollutants in ambient air around the sampling area.

  18. Chimeric RNase H–Competent Oligonucleotides Directed to the HIV-1 Rev Response Element

    PubMed Central

    Prater, Chrissy E.; Saleh, Anthony D.; Wear, Maggie P.; Miller, Paul S.

    2007-01-01

    Chimeric oligo-2′-O-methylribonucleotides containing centrally located patches of contiguous 2′-deoxyribonucleotides and terminating in a nuclease resistant 3′-methylphosphonate internucleotide linkage were prepared. The oligonucleotides were targeted to the 3′-side of HIV Rev response element (RRE) stem-loop IIB RNA, which is adjacent to the high affinity Rev protein binding site and is critical to virus function. Thermal denaturation experiments showed that chimeric oligonucleotides form very stable duplexes with a complementary single-stranded RNA, and gel electrophoretic mobility shift assays (EMSA) showed that they bind with high affinity and specificity to RRE stem-loop II RNA (KD approximately 200 nM). The chimeric oligonucleotides promote RNase H-mediated hydrolysis of RRE stem-loop II RNA and have half lives exceeding 24 h when incubated in cell culture medium containing 10% fetal calf serum. One of the chimeric oligonucleotides inhibited RRE mediated expression of chloramphenicol acetyl transferase (CAT) approximately 60% at a concentration of 300 nM in HEK 293T cells co-transfected with p-RRE/CAT and p-Rev mammalian expression vectors. PMID:17566743

  19. 5 CFR 293.101 - Purpose and scope.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ....101 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.101 Purpose and scope. (a) This subpart sets forth basic policies governing the creation, development, maintenance, processing, use...

  20. National Performance Benchmarks for Modern Screening Digital Mammography: Update from the Breast Cancer Surveillance Consortium.

    PubMed

    Lehman, Constance D; Arao, Robert F; Sprague, Brian L; Lee, Janie M; Buist, Diana S M; Kerlikowske, Karla; Henderson, Louise M; Onega, Tracy; Tosteson, Anna N A; Rauscher, Garth H; Miglioretti, Diana L

    2017-04-01

    Purpose To establish performance benchmarks for modern screening digital mammography and assess performance trends over time in U.S. community practice. Materials and Methods This HIPAA-compliant, institutional review board-approved study measured the performance of digital screening mammography interpreted by 359 radiologists across 95 facilities in six Breast Cancer Surveillance Consortium (BCSC) registries. The study included 1 682 504 digital screening mammograms performed between 2007 and 2013 in 792 808 women. Performance measures were calculated according to the American College of Radiology Breast Imaging Reporting and Data System, 5th edition, and were compared with published benchmarks by the BCSC, the National Mammography Database, and performance recommendations by expert opinion. Benchmarks were derived from the distribution of performance metrics across radiologists and were presented as 50th (median), 10th, 25th, 75th, and 90th percentiles, with graphic presentations using smoothed curves. Results Mean screening performance measures were as follows: abnormal interpretation rate (AIR), 11.6 (95% confidence interval [CI]: 11.5, 11.6); cancers detected per 1000 screens, or cancer detection rate (CDR), 5.1 (95% CI: 5.0, 5.2); sensitivity, 86.9% (95% CI: 86.3%, 87.6%); specificity, 88.9% (95% CI: 88.8%, 88.9%); false-negative rate per 1000 screens, 0.8 (95% CI: 0.7, 0.8); positive predictive value (PPV) 1, 4.4% (95% CI: 4.3%, 4.5%); PPV2, 25.6% (95% CI: 25.1%, 26.1%); PPV3, 28.6% (95% CI: 28.0%, 29.3%); cancers stage 0 or 1, 76.9%; minimal cancers, 57.7%; and node-negative invasive cancers, 79.4%. Recommended CDRs were achieved by 92.1% of radiologists in community practice, and 97.1% achieved recommended ranges for sensitivity. Only 59.0% of radiologists achieved recommended AIRs, and only 63.0% achieved recommended levels of specificity. Conclusion The majority of radiologists in the BCSC surpass cancer detection recommendations for screening mammography; however, AIRs continue to be higher than the recommended rate for almost half of radiologists interpreting screening mammograms. © RSNA, 2016 Online supplemental material is available for this article.

  1. Effects of tibolone on fibrinogen and antithrombin III: A systematic review and meta-analysis of controlled trials.

    PubMed

    Bała, Małgorzata; Sahebkar, Amirhossein; Ursoniu, Sorin; Serban, Maria-Corina; Undas, Anetta; Mikhailidis, Dimitri P; Lip, Gregory Y H; Rysz, Jacek; Banach, Maciej

    2017-10-01

    Tibolone is a synthetic steroid with estrogenic, androgenic and progestogenic activity, but the evidence regarding its effects on fibrinogen and antithrombin III (ATIII) has not been conclusive. We assessed the impact of tibolone on fibrinogen and ATIII through a systematic review and meta-analysis of available randomized controlled trials (RCTs). The search included PUBMED, Web of Science, Scopus, and Google Scholar (up to January 31st, 2016) to identify controlled clinical studies investigating the effects of oral tibolone treatment on fibrinogen and ATIII. Overall, the impact of tibolone on plasma fibrinogen concentrations was reported in 10 trials comprising 11 treatment arms. Meta-analysis did not suggest a significant reduction of fibrinogen levels following treatment with tibolone (WMD: -5.38%, 95% CI: -11.92, +1.16, p=0.107). This result was robust in the sensitivity analysis and not influenced after omitting each of the included studies from meta-analysis. When the studies were categorized according to the duration of treatment, there was no effect in the subsets of trials lasting either <12months (WMD: -7.64%, 95% CI: -16.58, +1.29, p=0.094) or ≥12months (WMD: -0.62%, 95% CI: -8.40, +7.17, p=0.876). With regard to ATIII, there was no change following treatment with tibolone (WMD: +0.74%, 95% CI: -1.44, +2.93, p=0.505) and this effect was robust in sensitivity analysis. There was no differential effect of tibolone on plasma ATIII concentrations in trials with either <12months (WMD: +2.26%, 95% CI: -3.14, +7.66, p=0.411) or≥12months (WMD: +0.06%, 95% CI: -1.16, +1.28, p=0.926) duration. Consistent with the results of subgroup analysis, meta-regression did not suggest any significant association between the changes in plasma concentrations of fibrinogen (slope: +0.40; 95% CI: -0.39, +1.19; p=0.317) and ATIII (slope: -0.17; 95% CI: -0.54, +0.20; p=0.374) with duration of treatment. In conclusion, meta-analysis did not suggest a significant reduction of fibrinogen and ATIII levels following treatment with tibolone. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. A selective antagonist reveals a potential role of G protein-coupled receptor 55 in platelet and endothelial cell function.

    PubMed

    Kargl, Julia; Brown, Andrew J; Andersen, Liisa; Dorn, Georg; Schicho, Rudolf; Waldhoer, Maria; Heinemann, Akos

    2013-07-01

    The G protein-coupled receptor 55 (GPR55) is a lysophosphatidylinositol (LPI) receptor that is also responsive to certain cannabinoids. Although GPR55 has been implicated in several (patho)physiologic functions, its role remains enigmatic owing mainly to the lack of selective GPR55 antagonists. Here we show that the compound CID16020046 ((4-[4-(3-hydroxyphenyl)-3-(4-methylphenyl)-6-oxo-1H,4H,5H,6H-pyrrolo[3,4-c]pyrazol-5-yl] benzoic acid) is a selective GPR55 antagonist. In yeast cells expressing human GPR55, CID16020046 antagonized agonist-induced receptor activation. In human embryonic kidney (HEK293) cells stably expressing human GPR55, the compound behaved as an antagonist on LPI-mediated Ca²⁺ release and extracellular signal-regulated kinases activation, but not in HEK293 cells expressing cannabinoid receptor 1 or 2 (CB₁ or CB₂). CID16020046 concentration dependently inhibited LPI-induced activation of nuclear factor of activated T-cells (NFAT), nuclear factor κ of activated B cells (NF-κB) and serum response element, translocation of NFAT and NF-κB, and GPR55 internalization. It reduced LPI-induced wound healing in primary human lung microvascular endothelial cells and reversed LPI-inhibited platelet aggregation, suggesting a novel role for GPR55 in platelet and endothelial cell function. CID16020046 is therefore a valuable tool to study GPR55-mediated mechanisms in primary cells and tissues.

  3. Development and Validation of a Novel Dual Luciferase Reporter Gene Assay to Quantify Ebola Virus VP24 Inhibition of IFN Signaling

    PubMed Central

    Frau, Aldo; Sgarbanti, Marco; Orsatti, Roberto

    2018-01-01

    The interferon (IFN) system is the first line of defense against viral infections. Evasion of IFN signaling by Ebola viral protein 24 (VP24) is a critical event in the pathogenesis of the infection and, hence, VP24 is a potential target for drug development. Since no drugs target VP24, the identification of molecules able to inhibit VP24, restoring and possibly enhancing the IFN response, is a goal of concern. Accordingly, we developed a dual signal firefly and Renilla luciferase cell-based drug screening assay able to quantify IFN-mediated induction of Interferon Stimulated Genes (ISGs) and its inhibition by VP24. Human Embryonic Kidney 293T (HEK293T) cells were transiently transfected with a luciferase reporter gene construct driven by the promoter of ISGs, Interferon-Stimulated Response Element (ISRE). Stimulation of cells with IFN-α activated the IFN cascade leading to the expression of ISRE. Cotransfection of cells with a plasmid expressing VP24 cloned from a virus isolated during the last 2014 outbreak led to the inhibition of ISRE transcription, quantified by a luminescent signal. To adapt this system to test a large number of compounds, we performed it in 96-well plates; optimized the assay analyzing different parameters; and validated the system by calculating the Z′- and Z-factor, which showed values of 0.62 and 0.53 for IFN-α stimulation assay and VP24 inhibition assay, respectively, indicative of robust assay performance. PMID:29495311

  4. CRISPR/Cas9 Mediated GFP Knock-in at the MAP1LC3B Locus in 293FT Cells Is Better for Bona Fide Monitoring Cellular Autophagy.

    PubMed

    Wu, Zhiqiang; Zhao, Jinlin; Qiu, Minghan; Mi, Zeyun; Meng, Maobin; Guo, Yu; Wang, Hui; Yuan, Zhiyong

    2018-04-19

    Accurately identifying and quantifying cellular autophagy is very important as the significance of autophagy in physiological and pathological processes becomes increasingly evident. Ectopically expressed fluorescent-tagged microtubule-associated protein light chain 3B (MAP1LC3B, LC3) is the most widely used reporter for monitoring autophagy activity thus far. However, this approach ignores the influence of constitutively overexpressed LC3 on autophagy itself and autophagy-related processes and its accuracy in indicating autophagy is questionable. Here, we generated a knock-in GFP-LC3 reporter via the CRISPR/Cas9 system in 293FT cells to add GFP to the N-terminal of and in frame with endogenous LC3. We proved that this knock-in GFP-LC3 was expressed at biological level driven by the endogenous transcriptional regulatory elements as the wild type alleles. Compared with the ectopically expressed GFP-LC3, the endogenous knock-in reporter exhibited much higher sensitivity and signal-to-noise ratio of GFP-LC3 puncta upon the induction or inhibition of autophagy at certain step for monitoring autophagy activity. Thus, according to the previous reported concerning and the results presented here, we suggest that this knock-in GFP-LC3 reporter is better for bona fide monitoring cellular autophagy and should be employed for further study of autophagy in vitro and in vivo. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Development and Validation of a Novel Dual Luciferase Reporter Gene Assay to Quantify Ebola Virus VP24 Inhibition of IFN Signaling.

    PubMed

    Fanunza, Elisa; Frau, Aldo; Sgarbanti, Marco; Orsatti, Roberto; Corona, Angela; Tramontano, Enzo

    2018-02-24

    The interferon (IFN) system is the first line of defense against viral infections. Evasion of IFN signaling by Ebola viral protein 24 (VP24) is a critical event in the pathogenesis of the infection and, hence, VP24 is a potential target for drug development. Since no drugs target VP24, the identification of molecules able to inhibit VP24, restoring and possibly enhancing the IFN response, is a goal of concern. Accordingly, we developed a dual signal firefly and Renilla luciferase cell-based drug screening assay able to quantify IFN-mediated induction of Interferon Stimulated Genes (ISGs) and its inhibition by VP24. Human Embryonic Kidney 293T (HEK293T) cells were transiently transfected with a luciferase reporter gene construct driven by the promoter of ISGs, Interferon-Stimulated Response Element (ISRE). Stimulation of cells with IFN-α activated the IFN cascade leading to the expression of ISRE. Cotransfection of cells with a plasmid expressing VP24 cloned from a virus isolated during the last 2014 outbreak led to the inhibition of ISRE transcription, quantified by a luminescent signal. To adapt this system to test a large number of compounds, we performed it in 96-well plates; optimized the assay analyzing different parameters; and validated the system by calculating the Z'- and Z-factor, which showed values of 0.62 and 0.53 for IFN-α stimulation assay and VP24 inhibition assay, respectively, indicative of robust assay performance.

  6. 14 CFR 293.10 - Exemption.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Secretary for Aviation and International Affairs will, by notice, issue and periodically update a list... REGULATIONS INTERNATIONAL PASSENGER TRANSPORTATION Exemption From Filing Tariffs § 293.10 Exemption. (a) Air... carriers of the Parties; (ii) Whether the country's Government has disapproved or deterred U.S. carrier...

  7. 14 CFR 293.10 - Exemption.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Secretary for Aviation and International Affairs will, by notice, issue and periodically update a list... REGULATIONS INTERNATIONAL PASSENGER TRANSPORTATION Exemption From Filing Tariffs § 293.10 Exemption. (a) Air... carriers of the Parties; (ii) Whether the country's Government has disapproved or deterred U.S. carrier...

  8. 14 CFR 293.10 - Exemption.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Secretary for Aviation and International Affairs will, by notice, issue and periodically update a list... REGULATIONS INTERNATIONAL PASSENGER TRANSPORTATION Exemption From Filing Tariffs § 293.10 Exemption. (a) Air... carriers of the Parties; (ii) Whether the country's Government has disapproved or deterred U.S. carrier...

  9. 14 CFR 293.10 - Exemption.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Secretary for Aviation and International Affairs will, by notice, issue and periodically update a list... REGULATIONS INTERNATIONAL PASSENGER TRANSPORTATION Exemption From Filing Tariffs § 293.10 Exemption. (a) Air... carriers of the Parties; (ii) Whether the country's Government has disapproved or deterred U.S. carrier...

  10. 5 CFR 293.108 - Rules of conduct.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Basic Policies on Maintenance of Personnel Records § 293.108 Rules of conduct. (a) Scope. These rules of conduct apply to all Office and agency employees responsible for creation, development, maintenance, processing...

  11. 5 CFR 293.404 - Retention schedule.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... acceptable performance from the date of the written advance notice of the proposed removal or reduction in....404 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.404 Retention schedule. (a)(1) Except as provided...

  12. Efflux transport of chrysin and apigenin sulfates in HEK293 cells overexpressing SULT1A3: The role of multidrug resistance-associated protein 4 (MRP4/ABCC4).

    PubMed

    Li, Wan; Sun, Hua; Zhang, Xingwang; Wang, Huan; Wu, Baojian

    2015-11-01

    Efflux transport is a critical determinant to the pharmacokinetics of sulfate conjugates. Here we aimed to establish SULT1A3 stably transfected HEK293 cells, and to determine the contributions of BCRP and MRP transporters to excretion of chrysin and apigenin sulfates. The cDNA of SULT1A3 was stably introduced into HEK293 cells using a lentiviral vector, generating a sulfonation active cell line (i.e., SULT293 cells). Identification of sulfate transporters was achieved through chemical inhibition (using chemical inhibitors) and biological inhibition (using short-hairpin RNAs (shRNAs)) methods. Sulfated metabolites were rapidly generated and excreted upon incubation of SULT293 cells with chrysin and apigenin. Ko143 (a selective BCRP inhibitor) did not show inhibitory effects on sulfate disposition, whereas the pan-MRP inhibitor MK-571 caused significant reductions (38.5-64.3%, p<0.001) in sulfate excretion and marked elevations (160-243%, p<0.05) in sulfate accumulation. Further, two efflux transporters (BCRP and MRP4) expressed in the cells were knocked-down by shRNA-mediated silencing. Neither sulfate excretion nor sulfate accumulation was altered in BCRP knocked-down cells as compared to scramble cells. By contrast, MRP4 knock-down led to moderate decreases (17.1-20.6%, p<0.05) in sulfate excretion and increases (125-135%, p<0.05) in sulfate accumulation. In conclusion, MRP4 was identified as an exporter for chrysin and apigenin sulfates. The SULT1A3 modified HEK293 cells were an appropriate tool to study SULT1A3-mediated sulfonation and to characterize BCRP/MRP4-mediated sulfate transport. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. New VMD2 gene mutations identified in patients affected by Best vitelliform macular dystrophy

    PubMed Central

    Marchant, D; Yu, K; Bigot, K; Roche, O; Germain, A; Bonneau, D; Drouin‐Garraud, V; Schorderet, D F; Munier, F; Schmidt, D; Neindre, P Le; Marsac, C; Menasche, M; Dufier, J L; Fischmeister, R; Hartzell, C; Abitbol, M

    2007-01-01

    Purpose The mutations responsible for Best vitelliform macular dystrophy (BVMD) are found in a gene called VMD2. The VMD2 gene encodes a transmembrane protein named bestrophin‐1 (hBest1) which is a Ca2+‐sensitive chloride channel. This study was performed to identify disease‐specific mutations in 27 patients with BVMD. Because this disease is characterised by an alteration in Cl− channel function, patch clamp analysis was used to test the hypothesis that one of the VMD2 mutated variants causes the disease. Methods Direct sequencing analysis of the 11 VMD2 exons was performed to detect new abnormal sequences. The mutant of hBest1 was expressed in HEK‐293 cells and the associated Cl− current was examined using whole‐cell patch clamp analysis. Results Six new VMD2 mutations were identified, located exclusively in exons four, six and eight. One of these mutations (Q293H) was particularly severe. Patch clamp analysis of human embryonic kidney cells expressing the Q293H mutant showed that this mutant channel is non‐functional. Furthermore, the Q293H mutant inhibited the function of wild‐type bestrophin‐1 channels in a dominant negative manner. Conclusions This study provides further support for the idea that mutations in VMD2 are a necessary factor for Best disease. However, because variable expressivity of VMD2 was observed in a family with the Q293H mutation, it is also clear that a disease‐linked mutation in VMD2 is not sufficient to produce BVMD. The finding that the Q293H mutant does not form functional channels in the membrane could be explained either by disruption of channel conductance or gating mechanisms or by improper trafficking of the protein to the plasma membrane. PMID:17287362

  14. Effect of total hydroalcholic extract of Nigella sativa and its n-hexane and ethyl acetate fractions on ACHN and GP-293 cell lines.

    PubMed

    Shahraki, Samira; Khajavirad, Abolfazl; Shafei, Mohammad Naser; Mahmoudi, Mahmoud; Tabasi, Nafisa Sadat

    2016-01-01

    Medicinal plants are noted for their many advantages including the ability to treat diseases such as cancer. In this study, we examined the antitumor effect of the medicinal plant Nigella sativa on the morphology, survival, and apoptosis of ACHN (human renal adenocarcinoma) and GP-293 (normal renal epithelial) cell lines. From a hydroalcoholic extract of N. sativa, n-hexane and ethyl acetate fractions were extracted. Cells were treated with various concentrations of total hydroalcholic extract and n-hexane and ethyl acetate fractions; cell viability, morphological changes, and apoptosis were then determined. Results were presented as mean ± standard error of the mean (SEM). One-way analysis of variance (ANOVA) was applied for the statistical analysis of the data. The total extract and the fractions in a dose- and time-dependent manner reduced the cell viability in ACHN with no effect on the GP-293 cell line. In addition, the total extract resulted in more morphological changes in the ACHN cells compared to the GP-293 cells. The effect of the total extract in inducing apoptosis after 48 hours in the ACHN cell line was greater than in GP-293. In addition, the effect of the two fractions was lower than the total extract at all used concentrations. Therefore, the effect of total extract and n-hexane and ethyl acetate fractions of N. sativa on cell viability and apoptosis in the ACHN cell line is greater than in the GP-293 cell line. However, the effect of the total extract is higher than either of the two fractions on their own.

  15. Broadband Gerchberg-Saxton algorithm for freeform diffractive spectral filter design.

    PubMed

    Vorndran, Shelby; Russo, Juan M; Wu, Yuechen; Pelaez, Silvana Ayala; Kostuk, Raymond K

    2015-11-30

    A multi-wavelength expansion of the Gerchberg-Saxton (GS) algorithm is developed to design and optimize a surface relief Diffractive Optical Element (DOE). The DOE simultaneously diffracts distinct wavelength bands into separate target regions. A description of the algorithm is provided, and parameters that affect filter performance are examined. Performance is based on the spectral power collected within specified regions on a receiver plane. The modified GS algorithm is used to design spectrum splitting optics for CdSe and Si photovoltaic (PV) cells. The DOE has average optical efficiency of 87.5% over the spectral bands of interest (400-710 nm and 710-1100 nm). Simulated PV conversion efficiency is 37.7%, which is 29.3% higher than the efficiency of the better performing PV cell without spectrum splitting optics.

  16. Structural study of quasi-one-dimensional vanadium pyroxene LiVSi{sub 2}O{sub 6} single crystals

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ishii, Yuto; Matsushita, Yoshitaka; Oda, Migaku

    Single crystals of quasi-one-dimensional vanadium pyroxene LiVSi{sub 2}O{sub 6} were synthesized and the crystal structures at 293 K and 113 K were studied using X-ray diffraction experiments. We found a structural phase transition from the room-temperature crystal structure with space group C2/c to a low-temperature structure with space group P2{sub 1}/c, resulting from a rotational displacement of SiO{sub 4} tetrahedra. The temperature dependence of magnetic susceptibility shows a broad maximum around 116 K, suggesting an opening of the Haldane gap expected for one-dimensional antiferromagnets with S=1. However, an antiferromagnetic long-range order was developed below 24 K, probably caused by amore » weak inter-chain magnetic coupling in the compound. - Graphical abstract: Low temperature crystal structure of LiVSi{sub 2}O{sub 6} and an orbital arrangement within the V-O zig-zag chain along the c-axis. - Highlights: • A low temperature structure of LiVSi{sub 2}O{sub 6} was determined by single crystal X-ray diffraction measurements. • The origin of the structural transition is a rotational displacement of SiO{sub 4} tetrahedra. • The uniform orbital overlap in the V-O zigzag chain makes the system a quasi one-dimensional antiferromagnet.« less

  17. Archaea and Bacteria Acclimate to High Total Ammonia in a Methanogenic Reactor Treating Swine Waste

    PubMed Central

    Parameswaran, Prathap; Rittmann, Bruce E.

    2016-01-01

    Inhibition by ammonium at concentrations above 1000 mgN/L is known to harm the methanogenesis phase of anaerobic digestion. We anaerobically digested swine waste and achieved steady state COD-removal efficiency of around 52% with no fatty-acid or H2 accumulation. As the anaerobic microbial community adapted to the gradual increase of total ammonia-N (NH3-N) from 890 ± 295 to 2040 ± 30 mg/L, the Bacterial and Archaeal communities became less diverse. Phylotypes most closely related to hydrogenotrophic Methanoculleus (36.4%) and Methanobrevibacter (11.6%), along with acetoclastic Methanosaeta (29.3%), became the most abundant Archaeal sequences during acclimation. This was accompanied by a sharp increase in the relative abundances of phylotypes most closely related to acetogens and fatty-acid producers (Clostridium, Coprococcus, and Sphaerochaeta) and syntrophic fatty-acid Bacteria (Syntrophomonas, Clostridium, Clostridiaceae species, and Cloacamonaceae species) that have metabolic capabilities for butyrate and propionate fermentation, as well as for reverse acetogenesis. Our results provide evidence countering a prevailing theory that acetoclastic methanogens are selectively inhibited when the total ammonia-N concentration is greater than ~1000 mgN/L. Instead, acetoclastic and hydrogenotrophic methanogens coexisted in the presence of total ammonia-N of ~2000 mgN/L by establishing syntrophic relationships with fatty-acid fermenters, as well as homoacetogens able to carry out forward and reverse acetogenesis. PMID:27725793

  18. Archaea and Bacteria Acclimate to High Total Ammonia in a Methanogenic Reactor Treating Swine Waste.

    PubMed

    Esquivel-Elizondo, Sofia; Parameswaran, Prathap; Delgado, Anca G; Maldonado, Juan; Rittmann, Bruce E; Krajmalnik-Brown, Rosa

    2016-01-01

    Inhibition by ammonium at concentrations above 1000 mgN/L is known to harm the methanogenesis phase of anaerobic digestion. We anaerobically digested swine waste and achieved steady state COD-removal efficiency of around 52% with no fatty-acid or H 2 accumulation. As the anaerobic microbial community adapted to the gradual increase of total ammonia-N (NH 3 -N) from 890 ± 295 to 2040 ± 30 mg/L, the Bacterial and Archaeal communities became less diverse. Phylotypes most closely related to hydrogenotrophic Methanoculleus (36.4%) and Methanobrevibacter (11.6%), along with acetoclastic Methanosaeta (29.3%), became the most abundant Archaeal sequences during acclimation. This was accompanied by a sharp increase in the relative abundances of phylotypes most closely related to acetogens and fatty-acid producers ( Clostridium , Coprococcus , and Sphaerochaeta ) and syntrophic fatty-acid Bacteria ( Syntrophomonas , Clostridium , Clostridiaceae species, and Cloacamonaceae species) that have metabolic capabilities for butyrate and propionate fermentation, as well as for reverse acetogenesis. Our results provide evidence countering a prevailing theory that acetoclastic methanogens are selectively inhibited when the total ammonia-N concentration is greater than ~1000 mgN/L. Instead, acetoclastic and hydrogenotrophic methanogens coexisted in the presence of total ammonia-N of ~2000 mgN/L by establishing syntrophic relationships with fatty-acid fermenters, as well as homoacetogens able to carry out forward and reverse acetogenesis.

  19. Drug-using male clients of female sex workers who report being paid for sex: HIV/STI, demographic and drug use correlates

    PubMed Central

    Wagner, Karla D.; Pitpitan, Eileen V.; Chavarin, Claudia V.; Magis-Rodriguez, Carlos; Patterson, Thomas L.

    2013-01-01

    Background Research has focused on male clients of female sex workers (FSWs) and their risk for HIV/STIs. Yet, it is unclear whether the commercial sex behaviors of these men are limited to paying for sex, or whether they may also be paid for sex themselves. Methods We analyzed interview data and HIV/STI test results from 170 drug-using male clients of FSWs in Tijuana, Mexico, to determine the extent to which these men report being paid for sex and the association with positive HIV/STI results. Results Over one-quarter of men reported having been paid for sex in the past four months. In a multivariate logistic regression model, reporting having been paid for sex was significantly associated with testing positive for any HIV/STI (Adjusted Odds Ratio [AdjOR] 3.53, 95% C.I. 1.33–9.35), being bisexual (AdjOR 15.59, 95% C.I. 4.81–50.53), injection drug use in the past four months (AdjOR 2.65, 95% C.I. 1.16–6.03), and cocaine use in the past four months (AdjOR 2.93, 95% C.I. 1.22–7.01). Conclusions Findings suggest that drug-using male clients of FSWs may be characterized by unique risk profiles that require tailored HIV prevention interventions. PMID:23863514

  20. The resistance patterns of normal ocular bacterial flora to 4 fluoroquinolone antibiotics.

    PubMed

    Park, Shin Hae; Lim, Jeong-A; Choi, Jun-Sub; Kim, Kyung-A; Joo, Choun-Ki

    2009-01-01

    The purposes of this study were to determine the normal ocular bacterial flora isolated from patients undergoing anterior segment surgery and to evaluate their in vitro susceptibility to ciprofloxacin, levofloxacin, gatifloxacin, and moxifloxacin. During January 2006 to December 2006, conjunctival swabs taken from 385 eyes were inoculated onto 5% blood agar plates. The isolated bacteria were classified by analysis of 16s ribosomal DNA sequencing. Disk diffusion testing was performed in accordance with Clinical and Laboratory Standards Institute Performance Standards. Three hundred sixty-three microorganisms were isolated in 291 samples from 385 eyes. Gram-positive species predominated (89.8%, 326 of the 363 isolates), and Staphylococcus epidermidis was the most frequently isolated organism, accounting for 60.6% (220 of the 363 isolates). For 293 gram-positive isolates, the prevalence rates of in vitro resistance to ciprofloxacin, levofloxacin, gatifloxacin, and moxifloxacin were 22.2% (65 isolates), 11.6% (34), 2.7% (8), and 5.1% (15), respectively. Two of the gram-negative isolates were resistant to only ciprofloxacin (5.4%, 2 of 37 isolates) and not to other fluoroquinolones. Of 62 ciprofloxacin-resistant, coagulase-negative staphylococci, 32 (51.6%) showed coresistance to levofloxacin. Seven organisms were resistant to all the fluoroquinolones. Fluoroquinolones have activity against normal aerobic flora of the ocular surface. Normal ocular flora, especially gram-positive species, has low resistance to the fourth-generation fluoroquinolones -- gatifloxacin and moxifloxacin.

  1. Acceptability of cancer chemoprevention trials: impact of the design

    PubMed Central

    Maisonneuve, Anne-Sophie; Huiart, Laetitia; Rabayrol, Laetitia; Horsman, Doug; Didelot, Remi; Sobol, Hagay; Eisinger, Francois

    2008-01-01

    Background: Chemoprevention could significantly reduce cancer burden. Assessment of efficacy and risk/benefit balance is at best achieved through randomized clinical trials. Methods: At a periodic health examination center 1463 adults were asked to complete a questionnaire about their willingness to be involved in different kinds of preventive clinical trials. Results: Among the 851 respondents (58.2%), 228 (26.8%) agreed to participate in a hypothetical chemoprevention trial aimed at reducing the incidence of lung cancer and 116 (29.3%) of 396 women agreed to a breast cancer chemoprevention trial. Randomization would not restrain participation (acceptability rate: 87.7% for lung cancer and 93.0% for breast cancer). In these volunteers, short-term trials (1 year) reached a high level of acceptability: 71.5% and 73.7% for lung and breast cancer prevention respectively. In contrast long-term trials (5 years or more) were far less acceptable: 9.2% for lung cancer (OR=7.7 CI95% 4.4-14.0) and 10.5 % for breast cancer (OR=6.9 CI95% 3.2-15.8). For lung cancer prevention, the route of administration impacts on acceptability with higher rate 53.1% for a pill vs. 7.9% for a spray (OR=6.7 CI95% 3.6-12.9). Conclusion: Overall healthy individuals are not keen to be involved in chemo-preventive trials, the design of which could however increase the acceptability rate. PMID:18769562

  2. High transmission Ni compound refractive lens for high energy X-rays

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Brancewicz, M., E-mail: brancew@spring8.or.jp; Itou, M.; Sakurai, Y.

    We present a new planar Ni compound refractive lens for high energy X-rays (116 keV). The lens is composed of identical plano-concave elements with longitudinal parabolic grooves manufactured by a punch technique. In order to increase the lens transmission, the thickness of the single lens at the parabolic groove vertex was reduced to less than 5 μm and the radius of curvature was reduced to about 20 μm. The small radius of curvature allowed us to reduce the number of single elements needed to get the focal length of 3 m to 54 single lenses. The gain parameter has beenmore » significantly improved compared to the previous lenses due to higher transmission, but the focused beam size and its gain are not as good as expected, mostly due to the aberrations caused by the lens shape imperfections.« less

  3. Translating infection control guidelines into practice: implementation process within a health care institution.

    PubMed

    Raveis, Victoria H; Conway, Laurie J; Uchida, Mayuko; Pogorzelska-Maziarz, Monika; Larson, Elaine L; Stone, Patricia W

    2014-04-01

    Health-care-associated infections (HAIs) remain a major patient safety problem even as policy and programmatic efforts designed to reduce HAIs have increased. Although information on implementing effective infection control (IC) efforts has steadily grown, knowledge gaps remain regarding the organizational elements that improve bedside practice and accommodate variations in clinical care settings. We conducted in-depth, semistructured interviews in 11 hospitals across the United States with a range of hospital personnel involved in IC (n = 116). We examined the collective nature of IC and the organizational elements that can enable disparate groups to work together to prevent HAIs. Our content analysis of participants' narratives yielded a rich description of the organizational process of implementing adherence to IC. Findings document the dynamic, fluid, interactional, and reactive nature of this process. Three themes emerged: implementing adherence efforts institution-wide, promoting an institutional culture to sustain adherence, and contending with opposition to the IC mandate.

  4. Inositol Pyrophosphate Profiling of Two HCT116 Cell Lines Uncovers Variation in InsP8 Levels

    PubMed Central

    Gu, Chunfang; Wilson, Miranda S. C.; Jessen, Henning J.; Saiardi, Adolfo; Shears, Stephen B.

    2016-01-01

    The HCT116 cell line, which has a pseudo-diploid karotype, is a popular model in the fields of cancer cell biology, intestinal immunity, and inflammation. In the current study, we describe two batches of diverged HCT116 cells, which we designate as HCT116NIH and HCT116UCL. Using both gel electrophoresis and HPLC, we show that HCT116UCL cells contain 6-fold higher levels of InsP8 than HCT116NIH cells. This observation is significant because InsP8 is one of a group of molecules collectively known as ‘inositol pyrophosphates’ (PP-InsPs)—highly ‘energetic’ and conserved regulators of cellular and organismal metabolism. Variability in the cellular levels of InsP8 within divergent HCT116 cell lines could have impacted the phenotypic data obtained in previous studies. This difference in InsP8 levels is more remarkable for being specific; levels of other inositol phosphates, and notably InsP6 and 5-InsP7, are very similar in both HCT116NIH and HCT116UCL lines. We also developed a new HPLC procedure to record 1-InsP7 levels directly (for the first time in any mammalian cell line); 1-InsP7 comprised <2% of total InsP7 in HCT116NIH and HCT116UCL lines. The elevated levels of InsP8 in the HCT116UCL lines were not due to an increase in expression of the PP-InsP kinases (IP6Ks and PPIP5Ks), nor to a decrease in the capacity to dephosphorylate InsP8. We discuss how the divergent PP-InsP profiles of the newly-designated HCT116NIH and HCT116UCL lines should be considered an important research opportunity: future studies using these two lines may uncover new features that regulate InsP8 turnover, and may also yield new directions for studying InsP8 function. PMID:27788189

  5. 5 CFR 293.203 - Review of Office or agency practices.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ....203 Section 293.203 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS... practices. Reviews of agency personnel management policies and practices will be conducted to insure... necessary. Office or agency officials who have knowledge of violations of these regulations shall take...

  6. 5 CFR 293.203 - Review of Office or agency practices.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ....203 Section 293.203 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS... practices. Reviews of agency personnel management policies and practices will be conducted to insure... necessary. Office or agency officials who have knowledge of violations of these regulations shall take...

  7. 5 CFR 293.203 - Review of Office or agency practices.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ....203 Section 293.203 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS... practices. Reviews of agency personnel management policies and practices will be conducted to insure... necessary. Office or agency officials who have knowledge of violations of these regulations shall take...

  8. 5 CFR 293.203 - Review of Office or agency practices.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ....203 Section 293.203 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS... practices. Reviews of agency personnel management policies and practices will be conducted to insure... necessary. Office or agency officials who have knowledge of violations of these regulations shall take...

  9. 36 CFR 293.8 - Permanent structures and commercial services.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 2 2010-07-01 2010-07-01 false Permanent structures and... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.8 Permanent structures and commercial services. Motels, summer homes, stores, resorts, organization camps, hunting and fishing lodges, electronic installations, and...

  10. 14 CFR 293.21 - Incorporation of contract terms by reference.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS INTERNATIONAL PASSENGER TRANSPORTATION Effect of Exemption § 293... scheduled transportation of passengers in foreign air transportation, provided that: (a) The notice...) are complied with, to the extent applicable; (b) In addition to other remedies at law, a carrier may...

  11. 14 CFR 293.21 - Incorporation of contract terms by reference.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS INTERNATIONAL PASSENGER TRANSPORTATION Effect of Exemption § 293... scheduled transportation of passengers in foreign air transportation, provided that: (a) The notice...) are complied with, to the extent applicable; (b) In addition to other remedies at law, a carrier may...

  12. 14 CFR 293.21 - Incorporation of contract terms by reference.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS INTERNATIONAL PASSENGER TRANSPORTATION Effect of Exemption § 293... scheduled transportation of passengers in foreign air transportation, provided that: (a) The notice...) are complied with, to the extent applicable; (b) In addition to other remedies at law, a carrier may...

  13. 14 CFR 293.21 - Incorporation of contract terms by reference.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS INTERNATIONAL PASSENGER TRANSPORTATION Effect of Exemption § 293... scheduled transportation of passengers in foreign air transportation, provided that: (a) The notice...) are complied with, to the extent applicable; (b) In addition to other remedies at law, a carrier may...

  14. 14 CFR 293.21 - Incorporation of contract terms by reference.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS INTERNATIONAL PASSENGER TRANSPORTATION Effect of Exemption § 293... scheduled transportation of passengers in foreign air transportation, provided that: (a) The notice...) are complied with, to the extent applicable; (b) In addition to other remedies at law, a carrier may...

  15. 36 CFR 293.10 - Jurisdiction over wildlife and fish.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 36 Parks, Forests, and Public Property 2 2012-07-01 2012-07-01 false Jurisdiction over wildlife... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.10 Jurisdiction over wildlife and fish. Nothing in the... States with respect to wildlife and fish in the National Forests. ...

  16. 36 CFR 293.10 - Jurisdiction over wildlife and fish.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 2 2011-07-01 2011-07-01 false Jurisdiction over wildlife... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.10 Jurisdiction over wildlife and fish. Nothing in the... States with respect to wildlife and fish in the National Forests. ...

  17. 36 CFR 293.10 - Jurisdiction over wildlife and fish.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 2 2010-07-01 2010-07-01 false Jurisdiction over wildlife... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.10 Jurisdiction over wildlife and fish. Nothing in the... States with respect to wildlife and fish in the National Forests. ...

  18. 36 CFR 293.10 - Jurisdiction over wildlife and fish.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 36 Parks, Forests, and Public Property 2 2013-07-01 2013-07-01 false Jurisdiction over wildlife... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.10 Jurisdiction over wildlife and fish. Nothing in the... States with respect to wildlife and fish in the National Forests. ...

  19. 36 CFR 293.10 - Jurisdiction over wildlife and fish.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 36 Parks, Forests, and Public Property 2 2014-07-01 2014-07-01 false Jurisdiction over wildlife... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.10 Jurisdiction over wildlife and fish. Nothing in the... States with respect to wildlife and fish in the National Forests. ...

  20. 5 CFR 293.203 - Review of Office or agency practices.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ....203 Section 293.203 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS... practices. Reviews of agency personnel management policies and practices will be conducted to insure... necessary. Office or agency officials who have knowledge of violations of these regulations shall take...

  1. 5 CFR 293.503 - Implementing instructions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... PERSONNEL RECORDS Employee Medical File System Records § 293.503 Implementing instructions. Agencies must... must— (a) Describe overall operation of the system within the agency including the designation of the agency official who will be responsible for overall system management. When the agency has a medical...

  2. 5 CFR 293.503 - Implementing instructions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... PERSONNEL RECORDS Employee Medical File System Records § 293.503 Implementing instructions. Agencies must... must— (a) Describe overall operation of the system within the agency including the designation of the agency official who will be responsible for overall system management. When the agency has a medical...

  3. 5 CFR 293.503 - Implementing instructions.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... PERSONNEL RECORDS Employee Medical File System Records § 293.503 Implementing instructions. Agencies must... must— (a) Describe overall operation of the system within the agency including the designation of the agency official who will be responsible for overall system management. When the agency has a medical...

  4. 5 CFR 293.503 - Implementing instructions.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... PERSONNEL RECORDS Employee Medical File System Records § 293.503 Implementing instructions. Agencies must... must— (a) Describe overall operation of the system within the agency including the designation of the agency official who will be responsible for overall system management. When the agency has a medical...

  5. 0{nu}{beta}{beta}-decay nuclear matrix elements with self-consistent short-range correlations

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Simkovic, Fedor; Bogoliubov Laboratory of Theoretical Physics, JINR, RU-141 980 Dubna, Moscow region; Department of Nuclear Physics, Comenius University, Mlynska dolina F1, SK-842 15 Bratislava

    A self-consistent calculation of nuclear matrix elements of the neutrinoless double-beta decays (0{nu}{beta}{beta}) of {sup 76}Ge, {sup 82}Se, {sup 96}Zr, {sup 100}Mo, {sup 116}Cd, {sup 128}Te, {sup 130}Te, and {sup 136}Xe is presented in the framework of the renormalized quasiparticle random phase approximation (RQRPA) and the standard QRPA. The pairing and residual interactions as well as the two-nucleon short-range correlations are for the first time derived from the same modern realistic nucleon-nucleon potentials, namely, from the charge-dependent Bonn potential (CD-Bonn) and the Argonne V18 potential. In a comparison with the traditional approach of using the Miller-Spencer Jastrow correlations, matrix elementsmore » for the 0{nu}{beta}{beta} decay are obtained that are larger in magnitude. We analyze the differences among various two-nucleon correlations including those of the unitary correlation operator method (UCOM) and quantify the uncertainties in the calculated 0{nu}{beta}{beta}-decay matrix elements.« less

  6. Riboflavin Depletion Promotes Tumorigenesis in HEK293T and NIH3T3 Cells by Sustaining Cell Proliferation and Regulating Cell Cycle-Related Gene Transcription.

    PubMed

    Long, Lin; He, Jian-Zhong; Chen, Ye; Xu, Xiu-E; Liao, Lian-Di; Xie, Yang-Min; Li, En-Min; Xu, Li-Yan

    2018-05-07

    Riboflavin is an essential component of the human diet and its derivative cofactors play an established role in oxidative metabolism. Riboflavin deficiency has been linked with various human diseases. The objective of this study was to identify whether riboflavin depletion promotes tumorigenesis. HEK293T and NIH3T3 cells were cultured in riboflavin-deficient or riboflavin-sufficient medium and passaged every 48 h. Cells were collected every 5 generations and plate colony formation assays were performed to observe cell proliferation. Subcutaneous tumorigenicity assays in NU/NU mice were used to observe tumorigenicity of riboflavin-depleted HEK293T cells. Mechanistically, gene expression profiling and gene ontology analysis were used to identify abnormally expressed genes induced by riboflavin depletion. Western blot analyses, cell cycle analyses, and chromatin immunoprecipitation were used to validate the expression of cell cycle-related genes. Plate colony formation of NIH3T3 and HEK293T cell lines was enhanced >2-fold when cultured in riboflavin-deficient medium for 10-20 generations. Moreover, we observed enhanced subcutaneous tumorigenicity in NU/NU mice following injection of riboflavin-depleted compared with normal HEK293T cells (55.6% compared with 0.0% tumor formation, respectively). Gene expression profiling and gene ontology analysis revealed that riboflavin depletion induced the expression of cell cycle-related genes. Validation experiments also found that riboflavin depletion decreased p21 and p27 protein levels by ∼20%, and increased cell cycle-related and expression-elevated protein in tumor (CREPT) protein expression >2-fold, resulting in cyclin D1 and CDK4 levels being increased ∼1.5-fold, and cell cycle acceleration. We also observed that riboflavin depletion decreased intracellular riboflavin levels by 20% and upregulated expression of riboflavin transporter genes, particularly SLC52A3, and that the changes in CREPT and SLC52A3 correlated with specific epigenetic changes in their promoters in riboflavin-depleted HEK293T cells. Riboflavin depletion contributes to HEK293T and NIH3T3 cell tumorigenesis and may be a risk factor for tumor development.

  7. Synthesis and Evaluation of a Novel 64Cu- and 67Ga-Labeled Neurokinin 1 Receptor Antagonist for in Vivo Targeting of NK1R-Positive Tumor Xenografts.

    PubMed

    Zhang, Hanwen; Kanduluru, Ananda Kumar; Desai, Pooja; Ahad, Afruja; Carlin, Sean; Tandon, Nidhi; Weber, Wolfgang A; Low, Philip S

    2018-04-18

    Neurokinin 1 receptor (NK1R) is expressed in gliomas and neuroendocrine malignancies and represents a promising target for molecular imaging and targeted radionuclide therapy. The goal of this study was to synthesize and evaluate a novel NK1R ligand (NK1R-NOTA) for targeting NK1R-expressing tumors. Using a carboxymethyl moiety linked to L-733060 as a starting reagent, NK1R-NOTA was synthesized in a three-step reaction and then labeled with 64 Cu (or 67 Ga for in vitro studies) in the presence of CH 3 COONH 4 buffer. The radioligand affinity and cellular uptake were evaluated with NK1R-transduced HEK293 cells (HEK293-NK1R) and NK1R nontransduced HEK293 cells (HEK293-WT) and their xenografts. Radiolabeled NK1R-NOTA was obtained with a radiochemical purity of >95% and specific activities of >7.0 GBq/μmol for 64 Cu and >5.0 GBq/μmol for 67 Ga. Both 64 Cu- and 67 Ga-labeled NK1R-NOTA demonstrated high levels of uptake in HEK293-NK1R cells, whereas co-incubation with an excess of NK1R ligand L-733060 reduced the level of uptake by 90%. Positron emission tomography (PET) imaging showed that [ 64 Cu]NK1R-NOTA had a accumulated rapidly in HEK293-NK1R xenografts and a 10-fold lower level of uptake in HEK293-WT xenografts. Radioactivity was cleared by gastrointestinal tract and urinary systems. Biodistribution studies confirmed that the tumor-to-organ ratios were ≥5 for all studied organs at 1 h p.i., except kidneys, liver, and intestine, and that the tumor-to-intestine and tumor-to-kidney ratios were also improved 4 and 20 h post-injection. [ 64 Cu]NK1R-NOTA is a promising ligand for PET imaging of NK1R-expressing tumor xenografts. Delayed imaging with [ 64 Cu]NK1R-NOTA improves image contrast because of the continuous clearance of radioactivity from normal organs.

  8. Cell-type specific effects of endocytosis inhibitors on 5-hydroxytryptamine(2A) receptor desensitization and resensitization reveal an arrestin-, GRK2-, and GRK5-independent mode of regulation in human embryonic kidney 293 cells.

    PubMed

    Gray, J A; Sheffler, D J; Bhatnagar, A; Woods, J A; Hufeisen, S J; Benovic, J L; Roth, B L

    2001-11-01

    The effect of endocytosis inhibitors on 5-hydroxytryptamine(2A) (5-HT(2A)) receptor desensitization and resensitization was examined in transiently transfected human embryonic kidney (HEK) 293 cells and in C6 glioma cells that endogenously express 5-HT(2A) receptors. In HEK-293 cells, 5-HT(2A) receptor desensitization was unaffected by cotransfection with a dominant-negative mutant of dynamin (DynK44A), a truncation mutant of arrestin-2 [Arr2(319-418)], or by two well-characterized chemical inhibitors of endocytosis: concanavalin A (conA) and phenylarsine oxide (PAO). In contrast, beta 2-adrenergic receptor desensitization was significantly potentiated by each of these treatments in HEK-293 cells. In C6 glioma cells, however, DynK44A, Arr2(319-418), conA, and PAO each resulted in the potentiation of 5-HT(2A) and beta-adrenergic receptor desensitization. The cell-type-specific effect of Arr2(319-418) on 5-HT(2A) receptor desensitization was not related to the level of GRK2 or GRK5 expression. Interestingly, although beta 2-adrenergic receptor resensitization was potently blocked by cotransfection with DynK44A, 5-HT(2A) receptor resensitization was enhanced, suggesting the existence of a novel cell-surface mechanism for 5-HT(2A) receptor resensitization in HEK-293 cells. In addition, Arr2(319-418) had no effect on 5-HT(2A) receptor resensitization in HEK-293 cells, although it attenuated the resensitization of the beta 2-adrenergic receptor. However, in C6 glioma cells, both DynK44A and Arr2(319-418) significantly reduced 5-HT(2A) receptor resensitization. Taken together, these results provide the first convincing evidence of cell-type-specific roles for endocytosis inhibitors in regulating GPCR activity. Additionally, these results imply that novel GRK and arrestin-independent mechanisms of 5-HT(2A) receptor desensitization and resensitization exist in HEK-293 cells.

  9. Search for double beta decay of 116Cd with enriched 116CdWO4 crystal scintillators (Aurora experiment)

    NASA Astrophysics Data System (ADS)

    Danevich, F. A.; Barabash, A. S.; Belli, P.; Bernabei, R.; Cappella, F.; Caracciolo, V.; Cerulli, R.; Chernyak, D. M.; d'Angelo, S.; Incicchitti, A.; Kobychev, V. V.; Konovalov, S. I.; Laubenstein, M.; Mokina, V. M.; Poda, D. V.; Polischuk, O. G.; Shlegel, V. N.; Tretyak, V. I.; Umatov, V. I.

    2016-05-01

    The Aurora experiment to investigate double beta decay of 116 Cd with the help of 1.162 kg cadmium tungstate crystal scintillators enriched in 116 Cd to 82% is in progress at the Gran Sasso Underground Laboratory. The half-life of 116 Cd relatively to the two neutrino double beta decay is measured with the highest up-to-date accuracy T1/2 = (2.62 ± 0.14) × 1019 yr. The sensitivity of the experiment to the neutrinoless double beta decay of 116 Cd to the ground state of 116 Sn is estimated as T1/2 ≥ 1.9 × 1023 yr at 90% CL, which corresponds to the effective Majorana neutrino mass limit (mv) ≤ (1.2 — 1.8) eV. New limits are obtained for the double beta decay of 116 Cd to the excited levels of 116 Sn, and for the neutrinoless double beta decay with emission of majorons.

  10. 5 CFR 293.102 - Definitions.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Policies on Maintenance of Personnel Records § 293.102 Definitions. In this part: Agency means any... President), or any independent regulatory agency; Data subject means the individual about whom the Office or agency is maintaining information in a system of records; Individual means a citizen of the United States...

  11. 5 CFR 293.102 - Definitions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Policies on Maintenance of Personnel Records § 293.102 Definitions. In this part: Agency means any... President), or any independent regulatory agency; Data subject means the individual about whom the Office or agency is maintaining information in a system of records; Individual means a citizen of the United States...

  12. 5 CFR 293.102 - Definitions.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Policies on Maintenance of Personnel Records § 293.102 Definitions. In this part: Agency means any... President), or any independent regulatory agency; Data subject means the individual about whom the Office or agency is maintaining information in a system of records; Individual means a citizen of the United States...

  13. 5 CFR 293.404 - Retention schedule.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... RECORDS Employee Performance File System Records § 293.404 Retention schedule. (a)(1) Except as provided... in this system, including transfer with the employee who changes positions, shall be as agencies... performance-related documents five years old or less shall be forwarded in the Employee Performance File along...

  14. 5 CFR 293.403 - Contents of employee performance files.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.403 Contents of employee performance files. (a) A decision on what constitutes a performance-related document within the meaning of... 5 Administrative Personnel 1 2011-01-01 2011-01-01 false Contents of employee performance files...

  15. 5 CFR 293.403 - Contents of employee performance files.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.403 Contents of employee performance files. (a) A decision on what constitutes a performance-related document within the meaning of... 5 Administrative Personnel 1 2013-01-01 2013-01-01 false Contents of employee performance files...

  16. 5 CFR 293.404 - Retention schedule.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... RECORDS Employee Performance File System Records § 293.404 Retention schedule. (a)(1) Except as provided... in this system, including transfer with the employee who changes positions, shall be as agencies... performance-related documents five years old or less shall be forwarded in the Employee Performance File along...

  17. 5 CFR 293.403 - Contents of employee performance files.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.403 Contents of employee performance files. (a) A decision on what constitutes a performance-related document within the meaning of... 5 Administrative Personnel 1 2014-01-01 2014-01-01 false Contents of employee performance files...

  18. 5 CFR 293.403 - Contents of employee performance files.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... REGULATIONS PERSONNEL RECORDS Employee Performance File System Records § 293.403 Contents of employee performance files. (a) A decision on what constitutes a performance-related document within the meaning of... 5 Administrative Personnel 1 2012-01-01 2012-01-01 false Contents of employee performance files...

  19. 5 CFR 293.404 - Retention schedule.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... RECORDS Employee Performance File System Records § 293.404 Retention schedule. (a)(1) Except as provided... in this system, including transfer with the employee who changes positions, shall be as agencies... performance-related documents five years old or less shall be forwarded in the Employee Performance File along...

  20. 5 CFR 293.404 - Retention schedule.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... RECORDS Employee Performance File System Records § 293.404 Retention schedule. (a)(1) Except as provided... in this system, including transfer with the employee who changes positions, shall be as agencies... performance-related documents five years old or less shall be forwarded in the Employee Performance File along...

  1. High level over-expression of different NCX isoforms in HEK293 cell lines and primary neuronal cultures is protective following oxygen glucose deprivation.

    PubMed

    Cross, Jane L; Boulos, Sherif; Shepherd, Kate L; Craig, Amanda J; Lee, Sharon; Bakker, Anthony J; Knuckey, Neville W; Meloni, Bruno P

    2012-07-01

    In this study we have assessed sodium-calcium exchanger (NCX) protein over-expression on cell viability in primary rat cortical neuronal and HEK293 cell cultures when subjected to oxygen-glucose deprivation (OGD). In cortical neuronal cultures, NCX2 and NCX3 over-expression was achieved using adenoviral vectors, and following OGD increased neuronal survival from ≈20% for control vector treated cultures to ≈80% for both NCX isoforms. In addition, we demonstrated that NCX2 and NCX3 over-expression in cortical neuronal cultures enables neurons to maintain intracellular calcium at significantly lower levels than control vector treated cultures when exposed to high (9mM) extracellular calcium challenge. Further assessment of NCX activity during OGD was performed using HEK293 cell lines generated to over-express NCX1, NCX2 or NCX3 isoforms. While it was shown that NCX isoform expression differed considerably in the different HEK293 cell lines, high levels of NCX over-expression was associated with increased resistance to OGD. Taken together, our findings show that high levels of NCX over-expression increases neuronal and HEK293 cell survival following OGD, improves calcium management in neuronal cultures and provides additional support for NCX as a therapeutic target to reduce ischemic brain injury. Copyright © 2012 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

  2. Arylsulfatase B Mediates the Sulfonation-Transport Interplay in Human Embryonic Kidney 293 Cells Overexpressing Sulfotransferase 1A3.

    PubMed

    Zhao, Mengjing; Wang, Shuai; Li, Feng; Dong, Dong; Wu, Baojian

    2016-09-01

    Elucidating the intricate relationships between metabolic and transport pathways contributes to improved predictions of in vivo drug disposition and drug-drug interactions. Here we reported that inhibited excretion of conjugative metabolites [i.e., hesperetin 3'-O-sulfate (H3'S) and hesperetin 7-O-sulfate (H7S)] by MK-571 led to reduced metabolism of hesperetin (a maximal 78% reduction) in human embryonic kidney 293 cells overexpressing sulfotransferase 1A3 (named SULT293 cells). The strong dependence of cellular sulfonation on the efflux transport of generated sulfated metabolites revealed an interplay of sulfonation metabolism with efflux transport (or sulfonation-transport interplay). Polymerase chain reaction (PCR) and Western blot analyses demonstrated that SULT293 cells expressed multiple sulfatases such as arylsulfatase A (ARSA), ARSB, and ARSC. Of these three desulfonation enzymes, only ARSB showed significant activities toward hesperetin sulfates. The intrinsic clearance values for the hydrolysis of H3'S and H7S were estimated at 0.6 and 0.5 μl/h/mg, respectively. Furthermore, knockdown of ARSB attenuated the regulatory effect of efflux transporter on cellular sulfonation, whereas overexpression of ABSB enhanced the transporter effect. Taken together, the results indicated that ARSB mediated the sulfonation-transport interplay in SULT293 cells. Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.

  3. Release of tissue inhibitor of metalloproteinase-2 from alginate microcapsule encapsulating genetically engineered cells

    PubMed Central

    Kim, Yeon Seong; Jeong, Young-II; Jin, Shu-Guang; Pei, Jian; Wen, Min; Kim, In-Young; Moon, Kyung-Sub; Jung, Tae-Young; Ryu, Hyang-Hwa; Jung, Shin

    2013-01-01

    Background In this study, 293T cells were genetically engineered to secrete tissue inhibitor of metalloproteinase-2 (TIMP2) and encapsulated into alginate microcapsules to continuously release TIMP2 protein. Methods The anti-invasive potential of the microcapsules was studied in vitro using brain tumor cells. The TIMP2 gene was transfected to 293T cells, and genetically engineered 293TIMP2 cells were encapsulated into alginate microcapsules. Release of TIMP2 protein was detected with Western blot analysis and the anti-invasive potential against U87MG cells was tested using gelatin zymography and a Matrigel assay. Results Cell viability within the alginate microcapsules was maintained at a cell density of 5 × 106. Because polycationic polymers are helpful for maintaining the mechanical strength of microcapsules with good cell viability, the alginate microcapsules were reinforced with chitosan (0.1% w/v). Expression of TIMP2 protein in cell lysates and secretion of TIMP2 into the conditioned medium was confirmed by Western blot analysis. Alginate microcapsules encapsulating 293TIMP2 cells released TIMP2 protein into the medium efficiently, where the TIMP2 protein participated in degradation of the matrix metalloproteinase-2 enzyme and inhibited invasion of U87MG cells. Conclusion Alginate microcapsules encapsulating 293TIMP2 cells are promising candidates for anti-invasive treatment of glioma. PMID:24231999

  4. 47 CFR 80.293 - Check bearings by authorized ship personnel.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 47 Telecommunication 5 2011-10-01 2011-10-01 false Check bearings by authorized ship personnel. 80... RADIO SERVICES STATIONS IN THE MARITIME SERVICES Equipment Authorization for Compulsory Ships § 80.293 Check bearings by authorized ship personnel. The requirement for calibration by check bearings is met if...

  5. 47 CFR 80.293 - Check bearings by authorized ship personnel.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 47 Telecommunication 5 2013-10-01 2013-10-01 false Check bearings by authorized ship personnel. 80... RADIO SERVICES STATIONS IN THE MARITIME SERVICES Equipment Authorization for Compulsory Ships § 80.293 Check bearings by authorized ship personnel. The requirement for calibration by check bearings is met if...

  6. 47 CFR 80.293 - Check bearings by authorized ship personnel.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 47 Telecommunication 5 2014-10-01 2014-10-01 false Check bearings by authorized ship personnel. 80... RADIO SERVICES STATIONS IN THE MARITIME SERVICES Equipment Authorization for Compulsory Ships § 80.293 Check bearings by authorized ship personnel. The requirement for calibration by check bearings is met if...

  7. 47 CFR 80.293 - Check bearings by authorized ship personnel.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 47 Telecommunication 5 2012-10-01 2012-10-01 false Check bearings by authorized ship personnel. 80... RADIO SERVICES STATIONS IN THE MARITIME SERVICES Equipment Authorization for Compulsory Ships § 80.293 Check bearings by authorized ship personnel. The requirement for calibration by check bearings is met if...

  8. 50 CFR 648.293 - Tilefish trip limits.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... forth as follows: § 648.293 Tilefish accountability measures. (a) If the ACL is exceeded, the amount of the ACL overage that cannot be directly attributed to IFQ allocation holders having exceeded their IFQ allocation will be deducted from the ACL in the following fishing year. All overages directly attributable to...

  9. 50 CFR 648.293 - Tilefish accountability measures.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Measures for the Tilefish Fishery § 648.293 Tilefish accountability measures. (a) If the ACL is exceeded, the amount of the ACL overage that cannot be directly attributed to IFQ allocation holders having exceeded their IFQ allocation will be deducted from the ACL in the following fishing year. All overages...

  10. 50 CFR 648.293 - Tilefish accountability measures.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Measures for the Tilefish Fishery § 648.293 Tilefish accountability measures. (a) If the ACL is exceeded, the amount of the ACL overage that cannot be directly attributed to IFQ allocation holders having exceeded their IFQ allocation will be deducted from the ACL in the following fishing year. All overages...

  11. 50 CFR 648.293 - Tilefish accountability measures.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Measures for the Tilefish Fishery § 648.293 Tilefish accountability measures. (a) If the ACL is exceeded, the amount of the ACL overage that cannot be directly attributed to IFQ allocation holders having exceeded their IFQ allocation will be deducted from the ACL in the following fishing year. All overages...

  12. 5 CFR 293.406 - Disclosure of records.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... RECORDS Employee Performance File System Records § 293.406 Disclosure of records. Disclosure as used here... of official duties. Disclosure of information from this file system shall be made only as permitted..., only under a routine use published by the Office for the system of records covering these records...

  13. 5 CFR 293.406 - Disclosure of records.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... RECORDS Employee Performance File System Records § 293.406 Disclosure of records. Disclosure as used here... of official duties. Disclosure of information from this file system shall be made only as permitted..., only under a routine use published by the Office for the system of records covering these records...

  14. 5 CFR 293.406 - Disclosure of records.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... RECORDS Employee Performance File System Records § 293.406 Disclosure of records. Disclosure as used here... of official duties. Disclosure of information from this file system shall be made only as permitted..., only under a routine use published by the Office for the system of records covering these records...

  15. 5 CFR 293.406 - Disclosure of records.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... RECORDS Employee Performance File System Records § 293.406 Disclosure of records. Disclosure as used here... of official duties. Disclosure of information from this file system shall be made only as permitted..., only under a routine use published by the Office for the system of records covering these records...

  16. 5 CFR 293.406 - Disclosure of records.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... RECORDS Employee Performance File System Records § 293.406 Disclosure of records. Disclosure as used here... of official duties. Disclosure of information from this file system shall be made only as permitted..., only under a routine use published by the Office for the system of records covering these records...

  17. 5 CFR 293.107 - Special safeguards for automated records.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... for automated records. (a) In addition to following the security requirements of § 293.106 of this... security safeguards for data about individuals in automated records, including input and output documents, reports, punched cards, magnetic tapes, disks, and on-line computer storage. The safeguards must be in...

  18. 5 CFR 293.107 - Special safeguards for automated records.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... for automated records. (a) In addition to following the security requirements of § 293.106 of this... security safeguards for data about individuals in automated records, including input and output documents, reports, punched cards, magnetic tapes, disks, and on-line computer storage. The safeguards must be in...

  19. 5 CFR 293.107 - Special safeguards for automated records.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... for automated records. (a) In addition to following the security requirements of § 293.106 of this... security safeguards for data about individuals in automated records, including input and output documents, reports, punched cards, magnetic tapes, disks, and on-line computer storage. The safeguards must be in...

  20. 5 CFR 293.107 - Special safeguards for automated records.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... for automated records. (a) In addition to following the security requirements of § 293.106 of this... security safeguards for data about individuals in automated records, including input and output documents, reports, punched cards, magnetic tapes, disks, and on-line computer storage. The safeguards must be in...

  1. Trace element concentrations in two subpopulations of lesser snow geese from Wrangel Island, Russia

    USGS Publications Warehouse

    Hui, A.; Takekawa, John Y.; Baranyuk, Vasily V.; Litvin, K.V.

    1998-01-01

    Lesser snow geese (Anser c. caerulescens) from the Wrangel Island, Russia breeding colony spend the winter in two widely separated areas: the northern subpopulation in southern British Columbia and northern Washington and the southern subpopulation in the Central Valley of California. We examined 19 trace elements in the eggs and livers of geese from these two subpopulations to examine whether geese from the different wintering areas have similar trace element burdens. Eggs collected at the breeding colony from geese of the southern subpopulation had slightly higher levels of manganese, an element that can cause neurological damage and behavioral changes in chicks, than geese of the northern subpopulation. Livers from adult geese collected on the two wintering areas showed significant differences in trace elements including copper, iron, magnesium, molybdenum, and zinc. Copper concentrations in the livers of geese from the southern subpopulation were much higher than those from the northern subpopulation (x¯ = 116 vs. 46 ppm; dry weight). Elevated levels of copper may induce anemia in birds. The differences in trace element concentrations of these two subpopulations may be related to farming practices in their wintering areas. Geese from the northern subpopulation feed in pastures and coastal marshes and migrate along the coast, but geese from the southern subpopulation feed predominantly in rice fields and migrate over farm land. Copper and manganese are major components of fertilizers and fungicides commonly applied during rice cultivation.

  2. Content of selected elements in Boletus badius fruiting bodies growing in extremely polluted wastes.

    PubMed

    Mleczek, Mirosław; Siwulski, Marek; Mikołajczak, Patrycja; Gąsecka, Monika; Sobieralski, Krzysztof; Szymańczyk, Mateusz; Goliński, Piotr

    2015-01-01

    The aim of the study was to analyse levels of 17 trace elements and 5 major minerals in 11 Boletus badius fruiting bodies able to grow in extremely polluted waste (flotation tailings) and polluted soil in southern Poland. The presented data widen the limited literature data about the abilities of wild-growing mushroom species to grow on heavily contaminated substrates. Content of elements in waste, soil and mushrooms was analysed by flame atomic absorption spectrometry (FAAS) and cold vapour atomic absorption spectrometry (CVAAS - Hg). The industrial areas differed greatly as regards the content of elements in flotation tailings and soil; therefore differences in Ag, Ba, Cd, Co, Fe, Mo, Ni, Pb, Ca, K, Mg, Na and P accumulation in mushrooms were observed. The highest contents of elements in mushrooms were observed for: As, Al, Cu and Zn (86 ± 28, 549 ± 116, 341 ± 59 and 506 ± 40 mg kg(-1) dry matter, respectively). Calculated bioconcentration factor (BCF) values were higher than 1 for Al (15.1-16.9), Fe (10.6-24.4) and Hg (10.2-16.4) only. The main value of the presented results is the fact that one of the common wild-growing mushroom species was able to grow on flotation tailings containing over 22 g kg(-1) of As and, additionally, effective accumulation of other elements was observed. In view of the high content of the majority of analysed elements in fruiting bodies, edible mushrooms from such polluted areas are nonconsumable.

  3. STS-116 Crew Portrait

    NASA Technical Reports Server (NTRS)

    2006-01-01

    This is the STS-116 Crew Portrait. Pictured on the front row from left to right are: William Oefelein, pilot; Joan Higginbotham, mission specialist; and Mark Polansky, commander. On the back row, left to right, are: Robert Curbeam, Nicholas Patrick, Sunita Williams, and the European Space Agency's Christer Fuglesang, all mission specialists. Williams joined Expedition 14 in progress to serve as flight engineer aboard the International Space Station (ISS). Launched aboard the Space Shuttle Discovery on December 9, 2006, the seven delivered two high profile Marshall Space Flight Center (MSFC') payloads: The Lab-On-A Chip Application Development Portable Test System (LOCAD-PTS) and the Water Delivery System, a vital component of the Station's Oxygen Generation System. The primary mission objective was to deliver and install the P5 truss element. The P5 installation was conducted during the first of three space walks, and involved use of both the shuttle and station's robotic arms. The remainder of the mission included a major reconfiguration and activation of the ISS electrical and thermal control systems, as well as delivery of Zvezda Service Module debris panels, which will increase ISS protection from potential impacts of micro-meteorites and orbital debris.

  4. Buckling Testing and Analysis of Honeycomb Sandwich Panel Arc Segments of a Full-Scale Fairing Barrel. Part 2; 6-Ply In-Autoclave Facesheets

    NASA Technical Reports Server (NTRS)

    Pineda, Evan J.; Meyers, David E.; Kosareo, Daniel N.; Zalewski, Bart F.; Dixon, Genevieve D.

    2013-01-01

    Four honeycomb sandwich panel types, representing 1/16th arc segments of a 10-m diameter barrel section of the Heavy Lift Launch Vehicle (HLLV), were manufactured and tested under the NASA Composites for Exploration program and the NASA Constellation Ares V program. Two configurations were chosen for the panels: 6-ply facesheets with 1.125 in. honeycomb core and 8-ply facesheets with 1.000 in. honeycomb core. Additionally, two separate carbon fiber/epoxy material systems were chosen for the facesheets: in-autoclave IM7/977-3 and out-of-autoclave T40-800b/5320-1. Smaller 3- by 5-ft panels were cut from the 1/16th barrel sections. These panels were tested under compressive loading at the NASA Langley Research Center (LaRC). Furthermore, linear eigenvalue and geometrically nonlinear finite element analyses were performed to predict the compressive response of each 3- by 5-ft panel. This manuscript summarizes the experimental and analytical modeling efforts pertaining to the panels composed of 6-ply, IM7/977-3 facesheets (referred to as Panels B-1 and B-2). To improve the robustness of the geometrically nonlinear finite element model, measured surface imperfections were included in the geometry of the model. Both the linear and nonlinear models yield good qualitative and quantitative predictions. Additionally, it was correctly predicted that the panel would fail in buckling prior to failing in strength. Furthermore, several imperfection studies were performed to investigate the influence of geometric imperfections, fiber angle misalignments, and three-dimensional (3-D) effects on the compressive response of the panel.

  5. Buckling Testing and Analysis of Honeycomb Sandwich Panel Arc Segments of a Full-Scale Fairing Barrel Part 1: 8-Ply In-Autoclave Facesheets. Part 1; 8-Ply In-Autoclave Facesheets

    NASA Technical Reports Server (NTRS)

    Myers, David E.; Pineda, Evan J.; Zalewski, Bart F.; Kosareo, Daniel N.; Kellas, Sotiris

    2013-01-01

    Four honeycomb sandwich panels, representing 1/16th arc segments of a 10-m diameter barrel section of the heavy lift launch vehicle, were manufactured under the NASA Composites for Exploration program and the NASA Space Launch Systems program. Two configurations were chosen for the panels: 6-ply facesheets with 1.125 in. honeycomb core and 8-ply facesheets with 1.000 in. honeycomb core. Additionally, two separate carbon fiber/epoxy material systems were chosen for the facesheets: inautoclave IM7/977-3 and out-of-autoclave T40-800b/5320-1. Smaller 3.00- by 5.00-ft panels were cut from the 1/16th barrel sections. These panels were tested under compressive loading at the NASA Langley Research Center. Furthermore, linear eigenvalue and geometrically nonlinear finite element analysis was performed to predict the compressive response of the 3.00- by 5.00-ft panels. This manuscript summarizes the experimental and analytical modeling efforts pertaining to the panel composed of 8-ply, IM7/977-3 facesheets (referred to Panel A). To improve the robustness of the geometrically nonlinear finite element model, measured surface imperfections were included in the geometry of the model. Both the linear and nonlinear models yield good qualitative and quantitative predictions. Additionally, it was predicted correctly that the panel would fail in buckling prior to failing in strength. Furthermore, several imperfection studies were performed to investigate the influence of geometric imperfections, fiber misalignments, and three-dimensional (3 D) effects on the compressive response of the panel.

  6. Establishing physico-chemical reference conditions in Mediterranean streams according to the European Water Framework Directive.

    PubMed

    Sánchez-Montoya, María del Mar; Arce, Maria Isabel; Vidal-Abarca, María Rosario; Suárez, María Luisa; Prat, Narcís; Gómez, Rosa

    2012-05-01

    Type-specific physico-chemical reference conditions are required for the assessment of ecological status in the Water Framework Directive context, similarly to the biological and hydro-morphological elements. This directive emphasises that natural variability of quality elements in high status (reference condition) needs to be quantified. Mediterranean streams often present a marked seasonal pattern in hydrological, biological and geochemical processes which could affect physico-chemical reference conditions. This study establishes general physico-chemical reference conditions (oxygenation, nutrient, salinity and acidification conditions) for different Mediterranean stream types. 116 potential reference sites located in 23 Mediterranean catchments in Spain were sampled in spring, summer and autumn in 2003. All sites were subjected to a screening method for the selection of reference sites in Mediterranean streams (Mediterranean Reference Criteria) and classified using a pre-established stream typology that establishes five different stream types (temporary streams, evaporite-calcareous at medium altitude, siliceous headwaters, calcareous headwaters and large watercourses). Reference conditions (reference value and reference threshold equivalents to high-good class boundary) were calculated using two different methods according to the availability of reference sites: the reference site 75th percentile approach of all reference sites and the 25th percentile of the population approach. The majority of the studied potential reference sites (76 out of 116) were selected as reference sites. Regarding type-specific reference conditions, only siliceous headwaters could be considered different from the rest of stream types because lower conductivity and pH. All reference stream types presented seasonal differences as regards some parameters, except for temporary streams due to the high natural variation of this stream type. For those parameters which presented seasonal differences in a specific stream type, the least restrictive values were proposed as reference conditions. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. 7 CFR 1150.116 - Nutrition education.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 9 2014-01-01 2013-01-01 true Nutrition education. 1150.116 Section 1150.116 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (MARKETING... Order Definitions § 1150.116 Nutrition education. Nutrition education means those activities intended to...

  8. 7 CFR 1150.116 - Nutrition education.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 9 2013-01-01 2013-01-01 false Nutrition education. 1150.116 Section 1150.116 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (MARKETING... Order Definitions § 1150.116 Nutrition education. Nutrition education means those activities intended to...

  9. 7 CFR 1150.116 - Nutrition education.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 9 2012-01-01 2012-01-01 false Nutrition education. 1150.116 Section 1150.116 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing... Order Definitions § 1150.116 Nutrition education. Nutrition education means those activities intended to...

  10. 7 CFR 1150.116 - Nutrition education.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 9 2011-01-01 2011-01-01 false Nutrition education. 1150.116 Section 1150.116 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing... Order Definitions § 1150.116 Nutrition education. Nutrition education means those activities intended to...

  11. Origin discrimination of defatted pork via trace elements profiling, stable isotope ratios analysis, and multivariate statistical techniques.

    PubMed

    Park, Yu Min; Lee, Cheong Mi; Hong, Joon Ho; Jamila, Nargis; Khan, Naeem; Jung, Jong-Hyun; Jung, Young-Chul; Kim, Kyong Su

    2018-09-01

    This study verified the origin of 346 defatted Korean and non-Korean pork samples via trace elements profiling, and C and N stable isotope ratios analysis. The analyzed elements were 6 Li, 7 Li, 10 B, 11 B, 51 V , 50 Cr, 52 Cr, 53 Cr, 55 Mn, 58 Ni, 60 Ni, 59 Co, 63 Cu, 65 Cu, 64 Zn, 66 Zn, 69 Ga, 71 Ga, 75 As, 82 Se, 84 Sr, 86 Sr, 87 Sr, 88 Sr, 85 Rb, 94 Mo, 95 Mo, 97 Mo, 107 Ag, 109 Ag, 110 Cd, 111 Cd, 113 Cd, 112 Cd, 114 Cd, 116 Cd, 133 Cs, 206 Pb, 207 Pb, and 208 Pb. Content (mg/kg) of 51 V (0.012), 50 Cr (0.882), 75 As (0.017), 85 Rb (57.7), and 87 Sr (46.3) were high in Korean pork samples whereas 6 Li, 7 Li, 59 Co, 55 Mn, 58 Ni, 84 Sr, 86 Sr, 88 Sr, 111 Cd, and 133 Cs were found higher in non-Korean samples. The results of discriminant analysis showed that the trace elements content and stable isotope ratios were significant for the discrimination of geographical origins with a perfect discrimination rate of 100%. Copyright © 2018 Elsevier Ltd. All rights reserved.

  12. Mapping cis-Regulatory Domains in the Human Genome UsingMulti-Species Conservation of Synteny

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ahituv, Nadav; Prabhakar, Shyam; Poulin, Francis

    2005-06-13

    Our inability to associate distant regulatory elements with the genes that they regulate has largely precluded their examination for sequence alterations contributing to human disease. One major obstacle is the large genomic space surrounding targeted genes in which such elements could potentially reside. In order to delineate gene regulatory boundaries we used whole-genome human-mouse-chicken (HMC) and human-mouse-frog (HMF) multiple alignments to compile conserved blocks of synteny (CBS), under the hypothesis that these blocks have been kept intact throughout evolution at least in part by the requirement of regulatory elements to stay linked to the genes that they regulate. A totalmore » of 2,116 and 1,942 CBS>200 kb were assembled for HMC and HMF respectively, encompassing 1.53 and 0.86 Gb of human sequence. To support the existence of complex long-range regulatory domains within these CBS we analyzed the prevalence and distribution of chromosomal aberrations leading to position effects (disruption of a genes regulatory environment), observing a clear bias not only for mapping onto CBS but also for longer CBS size. Our results provide a genome wide data set characterizing the regulatory domains of genes and the conserved regulatory elements within them.« less

  13. 14 CFR 1245.116 - Miscellaneous provisions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 5 2011-01-01 2010-01-01 true Miscellaneous provisions. 1245.116 Section 1245.116 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION PATENTS AND OTHER INTELLECTUAL PROPERTY RIGHTS Patent Waiver Regulations § 1245.116 Miscellaneous provisions. (a) Filing of...

  14. 14 CFR 1245.116 - Miscellaneous provisions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 5 2010-01-01 2010-01-01 false Miscellaneous provisions. 1245.116 Section 1245.116 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION PATENTS AND OTHER INTELLECTUAL PROPERTY RIGHTS Patent Waiver Regulations § 1245.116 Miscellaneous provisions. (a) Filing of...

  15. 40 CFR 116.1 - Applicability.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Applicability. 116.1 Section 116.1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS DESIGNATION OF HAZARDOUS SUBSTANCES § 116.1 Applicability. This regulation designates hazardous substances under section 311(b)(2)(A...

  16. 7 CFR 1942.116 - Economic feasibility requirements.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 13 2010-01-01 2009-01-01 true Economic feasibility requirements. 1942.116 Section 1942.116 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE... Facilities Projects § 1942.116 Economic feasibility requirements. All projects financed under this section...

  17. 7 CFR 1942.116 - Economic feasibility requirements.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 13 2011-01-01 2009-01-01 true Economic feasibility requirements. 1942.116 Section 1942.116 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE... Facilities Projects § 1942.116 Economic feasibility requirements. All projects financed under this section...

  18. 7 CFR 1942.116 - Economic feasibility requirements.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 13 2013-01-01 2013-01-01 false Economic feasibility requirements. 1942.116 Section 1942.116 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE... Facilities Projects § 1942.116 Economic feasibility requirements. All projects financed under this section...

  19. 7 CFR 1942.116 - Economic feasibility requirements.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 13 2012-01-01 2012-01-01 false Economic feasibility requirements. 1942.116 Section 1942.116 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE... Facilities Projects § 1942.116 Economic feasibility requirements. All projects financed under this section...

  20. 7 CFR 1942.116 - Economic feasibility requirements.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 13 2014-01-01 2013-01-01 true Economic feasibility requirements. 1942.116 Section 1942.116 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE... Facilities Projects § 1942.116 Economic feasibility requirements. All projects financed under this section...

  1. 40 CFR 116.1 - Applicability.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Applicability. 116.1 Section 116.1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS DESIGNATION OF HAZARDOUS SUBSTANCES § 116.1 Applicability. This regulation designates hazardous substances under section 311(b)(2)(A...

  2. 40 CFR 116.1 - Applicability.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 22 2011-07-01 2011-07-01 false Applicability. 116.1 Section 116.1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS DESIGNATION OF HAZARDOUS SUBSTANCES § 116.1 Applicability. This regulation designates hazardous substances under section 311(b)(2)(A...

  3. 30 CFR 250.293 - What operations require approval of the DWOP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... OFFSHORE OIL AND GAS AND SULPHUR OPERATIONS IN THE OUTER CONTINENTAL SHELF Plans and Information Deepwater Operations Plans (dwop) § 250.293 What operations require approval of the DWOP? You may not begin production... 30 Mineral Resources 2 2010-07-01 2010-07-01 false What operations require approval of the DWOP...

  4. 40 CFR 421.293 - Effluent limitations guidelines representing the degree of effluent reduction attainable by the...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Tin Subcategory § 421.293 Effluent limitations guidelines representing the degree of effluent... application of the best available technology economically achievable: (a) Tin smelter SO2 scrubber. BAT Limitations for the Secondary Tin Subcategory Pollutant or pollutant property Maximum for any 1 day Maximum...

  5. 40 CFR 421.293 - Effluent limitations guidelines representing the degree of effluent reduction attainable by the...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Tin Subcategory § 421.293 Effluent limitations guidelines representing the degree of effluent... application of the best available technology economically achievable: (a) Tin smelter SO2 scrubber. BAT Limitations for the Secondary Tin Subcategory Pollutant or pollutant property Maximum for any 1 day Maximum...

  6. 40 CFR 421.293 - Effluent limitations guidelines representing the degree of effluent reduction attainable by the...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Tin Subcategory § 421.293 Effluent limitations guidelines representing the degree of effluent... application of the best available technology economically achievable: (a) Tin smelter SO2 scrubber. BAT Limitations for the Secondary Tin Subcategory Pollutant or pollutant property Maximum for any 1 day Maximum...

  7. 40 CFR 421.293 - Effluent limitations guidelines representing the degree of effluent reduction attainable by the...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Tin Subcategory § 421.293 Effluent limitations guidelines representing the degree of effluent... application of the best available technology economically achievable: (a) Tin smelter SO2 scrubber. BAT Limitations for the Secondary Tin Subcategory Pollutant or pollutant property Maximum for any 1 day Maximum...

  8. 40 CFR 421.293 - Effluent limitations guidelines representing the degree of effluent reduction attainable by the...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Tin Subcategory § 421.293 Effluent limitations guidelines representing the degree of effluent... application of the best available technology economically achievable: (a) Tin smelter SO2 scrubber. BAT Limitations for the Secondary Tin Subcategory Pollutant or pollutant property Maximum for any 1 day Maximum...

  9. 5 CFR 293.303 - The roles and responsibilities of the Office, agencies, and custodians.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... contained within a document management system approved by the Office. (2) A custodian shall be responsible... MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.303 The roles and... such a position is part of the records of the Office of Personnel Management (Office). (b) The Office...

  10. 7 CFR 29.3 - Procedures for filing, hearing, and determination of applications.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... of Tobacco Inspection and Price Support Services to New Markets and to Additional Sales on Designated Markets § 29.3 Procedures for filing, hearing, and determination of applications. (a) Time and place of filing. Applications for the extension of tobacco inspection and price support services to new markets...

  11. 5 CFR 293.303 - The roles and responsibilities of the Office, agencies, and custodians.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... contained within a document management system approved by the Office. (2) A custodian shall be responsible... MANAGEMENT CIVIL SERVICE REGULATIONS PERSONNEL RECORDS Official Personnel Folder § 293.303 The roles and... such a position is part of the records of the Office of Personnel Management (Office). (b) The Office...

  12. 36 CFR 293.14 - Mineral leases and mineral permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 2 2011-07-01 2011-07-01 false Mineral leases and mineral... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.14 Mineral leases and mineral permits. (a) All laws pertaining to mineral leasing shall extend to each National Forest Wilderness for the period specified in the Wilderness...

  13. 36 CFR 293.14 - Mineral leases and mineral permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 2 2010-07-01 2010-07-01 false Mineral leases and mineral... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.14 Mineral leases and mineral permits. (a) All laws pertaining to mineral leasing shall extend to each National Forest Wilderness for the period specified in the Wilderness...

  14. 36 CFR 293.14 - Mineral leases and mineral permits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 36 Parks, Forests, and Public Property 2 2012-07-01 2012-07-01 false Mineral leases and mineral... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.14 Mineral leases and mineral permits. (a) All laws pertaining to mineral leasing shall extend to each National Forest Wilderness for the period specified in the Wilderness...

  15. 36 CFR 293.14 - Mineral leases and mineral permits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 36 Parks, Forests, and Public Property 2 2014-07-01 2014-07-01 false Mineral leases and mineral... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.14 Mineral leases and mineral permits. (a) All laws pertaining to mineral leasing shall extend to each National Forest Wilderness for the period specified in the Wilderness...

  16. 36 CFR 293.14 - Mineral leases and mineral permits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 36 Parks, Forests, and Public Property 2 2013-07-01 2013-07-01 false Mineral leases and mineral... AGRICULTURE WILDERNESS-PRIMITIVE AREAS § 293.14 Mineral leases and mineral permits. (a) All laws pertaining to mineral leasing shall extend to each National Forest Wilderness for the period specified in the Wilderness...

  17. Biochemical effects of veterinary antibiotics on proliferation and cell cycle arrest of human HEK293 cells.

    PubMed

    Kim, Hyeon Young; Kim, Ki-Tae; Kim, Sang Don

    2012-08-01

    The purpose of this study was to examine the effects of veterinary antibiotics, including amoxicillin (AMX), chlortetracycline (CTC) and tylosin (TYL), on the biochemical mechanism of human embryonic kidney cells (HEK293). CTC and TYL inhibited HEK293 cell proliferation, in both time- and dose-dependent manners, and changed the cell morphology; whereas, AMX showed no cytotoxic effects. The cell cycle analysis of CTC and TYL revealed G1-arrest in HEK293 cells. Western blot analysis also showed that CTC and TYL affected the activation of DNA damage responsive proteins, as well as cell cycle regulatory proteins, such as p53, p21(Waf1/Cip1) and Rb protein, which are crucial in the G1-S transition. The activation of p21(Waf1/Cip1) was significantly up-regulated over time, but there was no change in the level of CDK2 expression. The results of this study suggest that veterinary antibiotics, even at low level concentrations on continuous exposure, can potentially risk the development of human cells.

  18. Neurokinin 1 Receptor Mediates Membrane Blebbing and Sheer Stress-Induced Microparticle Formation in HEK293 Cells

    PubMed Central

    Chen, Panpan; Douglas, Steven D.; Meshki, John; Tuluc, Florin

    2012-01-01

    Cell-derived microparticles participate in intercellular communication similar to the classical messenger systems of small and macro-molecules that bind to specialized membrane receptors. Microparticles have been implicated in the regulation of a variety of complex physiopathologic processes, such as thrombosis, the control of innate and adaptive immunity, and cancer. The neurokinin 1 receptor (NK1R) is a Gq-coupled receptor present on the membrane of a variety of tissues, including neurons in the central and peripheral nervous system, immune cells, endocrine and exocrine glands, and smooth muscle. The endogenous agonist of NK1R is the undecapeptide substance P (SP). We have previously described intracellular signaling mechanisms that regulate NK1R-mediated rapid cell shape changes in HEK293 cells and U373MG cells. In the present study, we show that the activation of NK1R in HEK293 cells, but not in U373MG cells, leads to formation of sheer-stress induced microparticles that stain positive with the membrane-selective fluorescent dye FM 2–10. SP-induced microparticle formation is independent of elevated intracellular calcium concentrations and activation of NK1R present on HEK293-derived microparticles triggers detectable calcium increase in SP-induced microparticles. The ROCK inhibitor Y27632 and the dynamin inhibitor dynasore inhibited membrane blebbing and microparticle formation in HEK293 cells, strongly suggesting that microparticle formation in this cell type is dependent on membrane blebbing. PMID:23024816

  19. A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells

    PubMed Central

    Dietmair, Stefanie; Hodson, Mark P.; Quek, Lake-Ee; Timmins, Nicholas E.; Gray, Peter; Nielsen, Lars K.

    2012-01-01

    Hek293 cells are the predominant hosts for transient expression of recombinant proteins and are used for stable expression of proteins where post-translational modifications performed by CHO cells are inadequate. Nevertheless, there is little information available on the key cellular features underpinning recombinant protein production in Hek293 cells. To improve our understanding of recombinant protein production in Hek293 cells and identify targets for the engineering of an improved host cell line, we have compared a stable, recombinant protein producing Hek293 cell line and its parental cell line using a combination of transcriptomics, metabolomics and fluxomics. Producer cultures consumed less glucose than non-producer cultures while achieving the same growth rate, despite the additional burden of recombinant protein production. Surprisingly, there was no indication that producer cultures compensated for the reduction in glycolytic energy by increasing the efficiency of glucose utilization or increasing glutamine consumption. In contrast, glutamine consumption was lower and the majority of genes involved in oxidative phosphorylation were downregulated in producer cultures. We observed an overall downregulation of a large number of genes associated with broad cellular functions (e.g., cell growth and proliferation) in producer cultures, and therefore speculate that a broad adaptation of the cellular network freed up resources for recombinant protein production while maintaining the same growth rate. Increased abundance of genes associated with endoplasmic reticulum stress indicated a possible bottleneck at the point of protein folding and assembly. PMID:22937046

  20. Neurokinin 1 receptor mediates membrane blebbing and sheer stress-induced microparticle formation in HEK293 cells.

    PubMed

    Chen, Panpan; Douglas, Steven D; Meshki, John; Tuluc, Florin

    2012-01-01

    Cell-derived microparticles participate in intercellular communication similar to the classical messenger systems of small and macro-molecules that bind to specialized membrane receptors. Microparticles have been implicated in the regulation of a variety of complex physiopathologic processes, such as thrombosis, the control of innate and adaptive immunity, and cancer. The neurokinin 1 receptor (NK1R) is a Gq-coupled receptor present on the membrane of a variety of tissues, including neurons in the central and peripheral nervous system, immune cells, endocrine and exocrine glands, and smooth muscle. The endogenous agonist of NK1R is the undecapeptide substance P (SP). We have previously described intracellular signaling mechanisms that regulate NK1R-mediated rapid cell shape changes in HEK293 cells and U373MG cells. In the present study, we show that the activation of NK1R in HEK293 cells, but not in U373MG cells, leads to formation of sheer-stress induced microparticles that stain positive with the membrane-selective fluorescent dye FM 2-10. SP-induced microparticle formation is independent of elevated intracellular calcium concentrations and activation of NK1R present on HEK293-derived microparticles triggers detectable calcium increase in SP-induced microparticles. The ROCK inhibitor Y27632 and the dynamin inhibitor dynasore inhibited membrane blebbing and microparticle formation in HEK293 cells, strongly suggesting that microparticle formation in this cell type is dependent on membrane blebbing.

  1. PLK-1 Silencing in Bladder Cancer by siRNA Delivered With Exosomes.

    PubMed

    Greco, Kristin A; Franzen, Carrie A; Foreman, Kimberly E; Flanigan, Robert C; Kuo, Paul C; Gupta, Gopal N

    2016-05-01

    To use exosomes as a vector to deliver small interfering ribonucleic acid (siRNA) to silence the polo-like kinase 1 (PLK-1) gene in bladder cancer cells. Exosomes were isolated from both human embryonic kidney 293 (HEK293) cell and mesenchymal stem cell (MSC) conditioned media. Fluorescently labeled exosomes were co-cultured with bladder cancer and normal epithelial cells and uptake was quantified by image cytometry. PLK-1 siRNA and negative control siRNA were loaded into HEK293 and MSC exosomes using electroporation. An invasive bladder cancer cell line (UMUC3) was co-cultured with the electroporated exosomes. Quantitative reverse transcriptase polymerase chain reaction was performed. Protein analysis was performed by Western blot. Annexin V staining and MTT assays were used to investigate effects on apoptosis and viability. Bladder cancer cell lines internalize an increased percentage of HEK293 exosomes when compared to normal bladder epithelial cells. Treatment of UMUC3 cells with exosomes electroporated with PLK-1 siRNA achieved successful knockdown of PLK-1 mRNA and protein when compared to cells treated with negative control exosomes. HEK293 and MSC exosomes were effectively used as a delivery vector to transport PLK-1 siRNA to bladder cancer cells in vitro, resulting in selective gene silencing of PLK-1. The use of exosomes as a delivery vector for potential intravesical therapy is attractive. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. 40 CFR 428.116 - Pretreatment standards for new sources.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 30 2011-07-01 2011-07-01 false Pretreatment standards for new sources. 428.116 Section 428.116 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS RUBBER MANUFACTURING POINT SOURCE CATEGORY Latex Foam Subcategory § 428.116...

  3. 40 CFR 428.116 - Pretreatment standards for new sources.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 29 2010-07-01 2010-07-01 false Pretreatment standards for new sources. 428.116 Section 428.116 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS RUBBER MANUFACTURING POINT SOURCE CATEGORY Latex Foam Subcategory § 428.116...

  4. 32 CFR 536.116 - Responsibilities as to claims arising overseas under international agreements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... under international agreements. 536.116 Section 536.116 National Defense Department of Defense... Under International Agreements § 536.116 Responsibilities as to claims arising overseas under international agreements. (a) Command claims services or other responsible JA offices within whose jurisdiction...

  5. 32 CFR 536.116 - Responsibilities as to claims arising overseas under international agreements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... under international agreements. 536.116 Section 536.116 National Defense Department of Defense... Under International Agreements § 536.116 Responsibilities as to claims arising overseas under international agreements. (a) Command claims services or other responsible JA offices within whose jurisdiction...

  6. 32 CFR 536.116 - Responsibilities as to claims arising overseas under international agreements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... under international agreements. 536.116 Section 536.116 National Defense Department of Defense... Under International Agreements § 536.116 Responsibilities as to claims arising overseas under international agreements. (a) Command claims services or other responsible JA offices within whose jurisdiction...

  7. 32 CFR 536.116 - Responsibilities as to claims arising overseas under international agreements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... under international agreements. 536.116 Section 536.116 National Defense Department of Defense... Under International Agreements § 536.116 Responsibilities as to claims arising overseas under international agreements. (a) Command claims services or other responsible JA offices within whose jurisdiction...

  8. 32 CFR 536.116 - Responsibilities as to claims arising overseas under international agreements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... under international agreements. 536.116 Section 536.116 National Defense Department of Defense... Under International Agreements § 536.116 Responsibilities as to claims arising overseas under international agreements. (a) Command claims services or other responsible JA offices within whose jurisdiction...

  9. 42 CFR 438.116 - Solvency standards.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 42 Public Health 4 2010-10-01 2010-10-01 false Solvency standards. 438.116 Section 438.116 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL ASSISTANCE PROGRAMS MANAGED CARE Enrollee Rights and Protections § 438.116 Solvency standards. (a...

  10. 9 CFR 116.5 - Reports.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Reports. 116.5 Section 116.5 Animals..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS RECORDS AND REPORTS § 116.5 Reports. (a) When required by the Administrator, reports containing accurate and complete information concerning...

  11. 9 CFR 116.5 - Reports.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Reports. 116.5 Section 116.5 Animals..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS RECORDS AND REPORTS § 116.5 Reports. (a) When required by the Administrator, reports containing accurate and complete information concerning...

  12. 9 CFR 116.5 - Reports.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Reports. 116.5 Section 116.5 Animals..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS RECORDS AND REPORTS § 116.5 Reports. (a) When required by the Administrator, reports containing accurate and complete information concerning...

  13. 9 CFR 116.5 - Reports.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Reports. 116.5 Section 116.5 Animals..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS RECORDS AND REPORTS § 116.5 Reports. (a) When required by the Administrator, reports containing accurate and complete information concerning...

  14. Design, analysis, and testing of a flexure-based vibration-assisted polishing device

    NASA Astrophysics Data System (ADS)

    Gu, Yan; Zhou, Yan; Lin, Jieqiong; Lu, Mingming; Zhang, Chenglong; Chen, Xiuyuan

    2018-05-01

    A vibration-assisted polishing device (VAPD) composed of leaf-spring and right-circular flexure hinges is proposed with the aim of realizing vibration-assisted machining along elliptical trajectories. To design the structure, energy methods and the finite-element method are used to calculate the performance of the proposed VAPD. An improved bacterial foraging optimization algorithm is used to optimize the structural parameters. In addition, the performance of the VAPD is tested experimentally. The experimental results indicate that the maximum strokes of the two directional mechanisms operating along the Z1 and Z2 directions are 29.5 μm and 29.3 μm, respectively, and the maximum motion resolutions are 10.05 nm and 10.01 nm, respectively. The maximum working bandwidth is 1,879 Hz, and the device has a good step response.

  15. Synthesis and Characterization of a New Benzoindole Derivative with Apoptotic Activity Against Colon Cancer Cells.

    PubMed

    Hajiaghaalipour, Fatemeh; Faraj, Fadhil L; Bagheri, Elham; Ali, Hapipah M; Abdulla, Mahmood Ameen; Majid, Nazia A

    2017-01-01

    Colorectal cancer is the third most common form of cancer in both men and women around the world. The chemistry and biological study of heterocyclic compounds have been an interesting area for a long time in pharmaceutical and medicinal chemistry. A new synthetic compound, 2-(1,1-dimethyl-1H-benzo[e]indol-2-yl)-3-((2-hydroxyphenyl)amino) acrylaldehyde, abbreviated as DBID, was prepared through the reaction of 2-(diformylmethylidene)-1,1- dimethylbenzo[e]indole with 2-aminophenol. The chemical structure of the synthesized compound was characterized by 1H NMR, 13C NMR and APT-NMR spectroscopy and confirmed by elemental analysis (CHN). The compound was screened for the antiproliferation effect against colorectal cancer cell line, HCT 116 and its possible mechanism of action was elucidated. To determine the IC50 value, the MTT assay was used and its apoptosisinducing effect was investigated. DBID inhibited the proliferation of HCT 116 cells with an IC50 of 9.32 µg/ml and significantly increased the levels of caspase -8, -9 and -3/7 in the treated cells compared to untreated cells. Apoptosis features in HCT 116 cell was detected in treated cells by using the AO/PI staining that confirmed that the cells had undergone remarkable morphological changes in apoptotic bodies. Furthermore, this changes in expression of caspase -8, -9 and -3 were confirmed by gene and protein quantification using RT-PCR and western blot analysis, respectively. The current study showed that the DBID compound has demonstrated chemotherapeutic activity which was evidenced by significant increases in the expression and activation of caspase and exploit the apoptotic signaling pathways to trigger cancer cell death. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  16. Osteopontin plays a pivotal role in increasing severity of respiratory syncytial virus infection

    PubMed Central

    Sampayo-Escobar, Viviana; Green, Ryan; Cheung, Michael B.; Bedi, Raminder; Mohapatra, Subhra

    2018-01-01

    The molecular mechanisms underlying susceptibility to severe respiratory syncytial virus (RSV) infection remain poorly understood. Herein, we report on the role of osteopontin (OPN) in regulation of RSV infection in human epithelial cells and how interleukin-1 beta (IL-1β), a cytokine secreted soon after RSV infection, when persistently expressed can induce OPN expression leading to increased viral infection. We first compared OPN expression in two human epithelial cell lines: HEK-293 and HEp-2. In contrast to HEp-2, HEK-293 expresses low levels of pro-caspase-1 resulting in decreased IL-1β expression in response to RSV infection. We found a correlation between low IL-1β levels and a delay in induction of OPN expression in RSV-infected HEK-293 cells compared to HEp-2. This phenomenon could partially explain the high susceptibility of HEp-2 cells to RSV infection versus the moderate susceptibility of HEK-293 cells. Also, HEK-293 cells expressing low levels of pro-caspase-1 exhibit decreased IL-1β expression and delayed OPN expression in response to RSV infection. HEK-293 cells incubated with human rIL-1β showed a dose-dependent increase in OPN expression upon RSV infection. Also, incubation with rOPN increased RSV viral load. Moreover, HEp-2 cells or mice infected with a mucogenic RSV strain RSV-L19F showed elevated levels of OPN in contrast to mice infected with the laboratory RSV strain rA2. This correlated with elevated levels of OPN following infection with RSV-L19F compared to rA2. Together, these results demonstrate that increased OPN expression is regulated in part by IL-1β, and the interplay between IL-1β and OPN signaling may play a pivotal role in the spread of RSV infection. PMID:29677209

  17. Genetic engineering of somatic cells to study and improve cardiac function.

    PubMed

    Kirkton, Robert D; Bursac, Nenad

    2012-11-01

    To demonstrate the utility of genetically engineered excitable cells for studies of basic electrophysiology and cardiac cell therapy. 'Zig-zag' networks of neonatal rat ventricular myocytes (NRVMs) were micropatterned onto thin elastomeric films to mimic the slow action potential (AP) conduction found in fibrotic myocardium. Addition of genetically engineered excitable human embryonic kidney cells (HEK-293 cells) ('Ex-293' cells stably expressing Kir2.1, Na(v)1.5, and Cx43 channels) increased both cardiac conduction velocity by 370% and twitch force amplitude by 64%. Furthermore, we stably expressed mutant Na(v)1.5 [A1924T (fast sodium channel mutant (substitution of alanine by threonine at amino acid 1924)] channels with hyperpolarized steady-state activation and showed that, despite a 71.6% reduction in peak I(Na), these cells propagated APs at the same velocity as the wild-type Na(v)1.5-expressing Ex-293 cells. Stable expression of Ca(v)3.3 (T-type voltage-gated calcium) channels in Ex-293 cells (to generate an 'ExCa-293' line) significantly increased their AP duration and reduced repolarization gradients in cocultures of these cells and NRVMs. Additional expression of an optogenetic construct [ChIEF (light-gated Channelrhodopsin mutant)]enabled light-based control of AP firing in ExCa-293 cells. We show that, despite being non-contractile, genetically engineered excitable cells can significantly improve both electrical and mechanical function of engineered cardiac tissues in vitro. We further demonstrate the utility of engineered cells for tissue-level studies of basic electrophysiology and cardiac channelopathies. In the future, this novel platform could be utilized in the high-throughput design of new genetically encoded indicators of cell electrical function, validation, and improvement of computer models of AP conduction, and development of novel engineered somatic cell therapies for the treatment of cardiac infarction and arrhythmias.

  18. Targeted repression of AXIN2 and MYC gene expression using designer TALEs

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rennoll, Sherri A.; Scott, Samantha A.; Yochum, Gregory S., E-mail: gsy3@psu.edu

    Highlights: • We designed TALE–SID fusion proteins to target AXIN2 and MYC. • TALE–SIDs bound the chromosomal AXIN2 and MYC genes and repressed their expression. • TALE–SIDs repress β-catenin{sup S45F}-dependent AXIN2 and MYC transcription. - Abstract: Designer TALEs (dTALEs) are chimeric transcription factors that can be engineered to regulate gene expression in mammalian cells. Whether dTALEs can block gene transcription downstream of signal transduction cascades, however, has yet to be fully explored. Here we tested whether dTALEs can be used to target genes whose expression is controlled by Wnt/β-catenin signaling. TALE DNA binding domains were engineered to recognize sequences adjacentmore » to Wnt responsive enhancer elements (WREs) that control expression of axis inhibition protein 2 (AXIN2) and c-MYC (MYC). These custom DNA binding domains were linked to the mSin3A interaction domain (SID) to generate TALE–SID chimeric repressors. The TALE–SIDs repressed luciferase reporter activity, bound their genomic target sites, and repressed AXIN2 and MYC expression in HEK293 cells. We generated a novel HEK293 cell line to determine whether the TALE–SIDs could function downstream of oncogenic Wnt/β-catenin signaling. Treating these cells with doxycycline and tamoxifen stimulates nuclear accumulation of a stabilized form of β-catenin found in a subset of colorectal cancers. The TALE–SIDs repressed AXIN2 and MYC expression in these cells, which suggests that dTALEs could offer an effective therapeutic strategy for the treatment of colorectal cancer.« less

  19. Transcriptional regulation of the novel monoamine oxidase renalase: Crucial roles of transcription factors Sp1, STAT3, and ZBP89.

    PubMed

    Sonawane, Parshuram J; Gupta, Vinayak; Sasi, Binu K; Kalyani, Ananthamohan; Natarajan, Bhargavi; Khan, Abrar A; Sahu, Bhavani S; Mahapatra, Nitish R

    2014-11-11

    Renalase, a novel monoamine oxidase, is emerging as an important regulator of cardiovascular, metabolic, and renal diseases. However, the mechanism of transcriptional regulation of this enzyme remains largely unknown. We undertook a systematic analysis of the renalase gene to identify regulatory promoter elements and transcription factors. Computational analysis coupled with transfection of human renalase promoter/luciferase reporter plasmids (5'-promoter-deletion constructs) into various cell types (HEK-293, IMR32, and HepG2) identified two crucial promoter domains at base pairs -485 to -399 and -252 to -150. Electrophoretic mobility shift assays using renalase promoter oligonucleotides with and without potential binding sites for transcription factors Sp1, STAT3, and ZBP89 displayed formation of specific complexes with HEK-293 nuclear proteins. Consistently, overexpression of Sp1, STAT3, and ZBP89 augmented renalase promoter activity; additionally, siRNA-mediated downregulation of Sp1, STAT3, and ZBP89 reduced the level of endogenous renalase transcription as well as the transfected renalase promoter activity. In addition, chromatin immunoprecipitation assays showed in vivo interactions of these transcription factors with renalase promoter. Interestingly, renalase promoter activity was augmented by nicotine and catecholamines; while Sp1 and STAT3 synergistically activated the nicotine-induced effect, Sp1 appeared to enhance epinephrine-evoked renalase transcription. Moreover, renalase transcript levels in mouse models of human essential hypertension were concomitantly associated with endogenous STAT3 and ZBP89 levels, suggesting crucial roles for these transcription factors in regulating renalase gene expression in cardiovascular pathological conditions.

  20. Protein kinase C activation potentiates gating of the vanilloid receptor VR1 by capsaicin, protons, heat and anandamide

    PubMed Central

    Vellani, Vittorio; Mapplebeck, Sarah; Moriondo, Andrea; Davis, John B; McNaughton, Peter A

    2001-01-01

    The effects of activation of protein kinase C (PKC) on membrane currents gated by capsaicin, protons, heat and anandamide were investigated in primary sensory neurones from neonatal rat dorsal root ganglia (DRG) and in HEK293 cells (human embryonic kidney cell line) transiently or stably expressing the human vanilloid receptor hVR1. Maximal activation of PKC by a brief application of phorbol 12-myristate 13-acetate (PMA) increased the mean membrane current activated by a low concentration of capsaicin by 1.65-fold in DRG neurones and 2.18-fold in stably transfected HEK293 cells. Bradykinin, which activates PKC, also enhanced the response to capsaicin in DRG neurones. The specific PKC inhibitor RO31-8220 prevented the enhancement caused by PMA. Activation of PKC did not enhance the membrane current at high concentrations of capsaicin, showing that PKC activation increases the probability of channel opening rather than unmasking channels. Application of PMA alone activated an inward current in HEK293 cells transiently transfected with VR1. The current was suppressed by the VR1 antagonist capsazepine. PMA did not, however, activate a current in the large majority of DRG neurones nor in HEK293 cells stably transfected with VR1. Removing external Ca2+ enhanced the response to a low concentration of capsaicin 2.40-fold in DRG neurones and 3.42-fold in HEK293 cells. Activation of PKC in zero Ca2+ produced no further enhancement of the response to capsaicin in either DRG neurones or HEK293 cells stably transfected with VR1. The effects of PKC activation on the membrane current gated by heat, anandamide and low pH were qualitatively similar to those on the capsaicin-gated current. The absence of a current activated by PMA in most DRG neurones or in stably transfected HEK293 cells suggests that activation of PKC does not directly open VR1 channels, but instead increases the probability that they will be activated by capsaicin, heat, low pH or anandamide. Removal of calcium also potentiates activation, and PKC activation then has no further effect. The results are consistent with a model in which phosphorylation of VR1 by PKC increases the probability of channel gating by agonists, and in which dephosphorylation occurs by a calcium-dependent process. PMID:11483711

  1. Vector modifications to eliminate transposase expression following piggyBac-mediated transgenesis

    PubMed Central

    Chakraborty, Syandan; Ji, HaYeun; Chen, Jack; Gersbach, Charles A.; Leong, Kam W.

    2014-01-01

    Transgene insertion plays an important role in gene therapy and in biological studies. Transposon-based systems that integrate transgenes by transposase-catalyzed “cut-and-paste” mechanism have emerged as an attractive system for transgenesis. Hyperactive piggyBac transposon is particularly promising due to its ability to integrate large transgenes with high efficiency. However, prolonged expression of transposase can become a potential source of genotoxic effects due to uncontrolled transposition of the integrated transgene from one chromosomal locus to another. In this study we propose a vector design to decrease post-transposition expression of transposase and to eliminate the cells that have residual transposase expression. We design a single plasmid construct that combines the transposase and the transpositioning transgene element to share a single polyA sequence for termination. Consequently, the separation of the transposase element from the polyA sequence after transposition leads to its deactivation. We also co-express Herpes Simplex Virus thymidine kinase (HSV-tk) with the transposase. Therefore, cells having residual transposase expression can be eliminated by the administration of ganciclovir. We demonstrate the utility of this combination transposon system by integrating and expressing a model therapeutic gene, human coagulation Factor IX, in HEK293T cells. PMID:25492703

  2. 32 CFR 219.116 - General requirements for informed consent.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 2 2014-07-01 2014-07-01 false General requirements for informed consent. 219.116 Section 219.116 National Defense Department of Defense (Continued) OFFICE OF THE SECRETARY OF DEFENSE (CONTINUED) MISCELLANEOUS PROTECTION OF HUMAN SUBJECTS § 219.116 General requirements for informed...

  3. 40 CFR 89.116 - Engine families.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 20 2011-07-01 2011-07-01 false Engine families. 89.116 Section 89.116... Provisions § 89.116 Engine families. (a) A manufacturer's product line is divided into engine families that... life periods. (b) The following characteristics distinguish engine families: (1) Fuel; (2) Cooling...

  4. 40 CFR 116.2 - Abbreviations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Abbreviations. 116.2 Section 116.2 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS DESIGNATION OF HAZARDOUS SUBSTANCES § 116.2 Abbreviations. ppm=parts per million mg=milligram(s) kg=kilogram(s) mg/l=milligrams(s) per...

  5. 7 CFR 1.16 - Extension of administrative deadlines.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 1 2013-01-01 2013-01-01 false Extension of administrative deadlines. 1.16 Section 1.16 Agriculture Office of the Secretary of Agriculture ADMINISTRATIVE REGULATIONS Official Records § 1.16 Extension of administrative deadlines. (a) In unusual circumstances as specified in this section...

  6. 40 CFR 405.116 - Pretreatment standards for new sources.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for new sources. 405.116 Section 405.116 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS DAIRY PRODUCTS PROCESSING POINT SOURCE CATEGORY Condensed Whey Subcategory § 405.116...

  7. 40 CFR 81.116 - Northern Missouri Intrastate Air Quality Control Region.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Quality Control Region. 81.116 Section 81.116 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Air Quality Control Regions § 81.116 Northern Missouri Intrastate Air Quality Control Region. The Northern Missouri Intrastate Air Quality Control Region consists of the territorial area encompassed by the...

  8. 40 CFR 81.116 - Northern Missouri Intrastate Air Quality Control Region.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Quality Control Region. 81.116 Section 81.116 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Air Quality Control Regions § 81.116 Northern Missouri Intrastate Air Quality Control Region. The Northern Missouri Intrastate Air Quality Control Region consists of the territorial area encompassed by the...

  9. 45 CFR 400.116 - Service for unaccompanied minors.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 45 Public Welfare 2 2012-10-01 2012-10-01 false Service for unaccompanied minors. 400.116 Section 400.116 Public Welfare Regulations Relating to Public Welfare OFFICE OF REFUGEE RESETTLEMENT... Child Welfare Services § 400.116 Service for unaccompanied minors. (a) A State must provide...

  10. 46 CFR 116.510 - Embarkation stations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Embarkation stations. 116.510 Section 116.510 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150... and Embarkation Station Requirements § 116.510 Embarkation stations. (a) A vessel must have a least...

  11. 46 CFR 116.510 - Embarkation stations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Embarkation stations. 116.510 Section 116.510 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150... and Embarkation Station Requirements § 116.510 Embarkation stations. (a) A vessel must have a least...

  12. 40 CFR 116.2 - Abbreviations.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Abbreviations. 116.2 Section 116.2 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS DESIGNATION OF HAZARDOUS SUBSTANCES § 116.2 Abbreviations. ppm=parts per million mg=milligram(s) kg=kilogram(s) mg/l=milligrams(s) per...

  13. 40 CFR 116.2 - Abbreviations.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 22 2011-07-01 2011-07-01 false Abbreviations. 116.2 Section 116.2 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS DESIGNATION OF HAZARDOUS SUBSTANCES § 116.2 Abbreviations. ppm=parts per million mg=milligram(s) kg=kilogram(s) mg/l=milligrams(s) per...

  14. 40 CFR 89.116 - Engine families.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Engine families. 89.116 Section 89.116... Provisions § 89.116 Engine families. (a) A manufacturer's product line is divided into engine families that... life periods. (b) The following characteristics distinguish engine families: (1) Fuel; (2) Cooling...

  15. 46 CFR 116.520 - Emergency evacuation plan.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Emergency evacuation plan. 116.520 Section 116.520... ARRANGEMENT Escape and Embarkation Station Requirements § 116.520 Emergency evacuation plan. The owner or managing operator shall prepare an evacuation plan that must: (a) Identify possible casualties involving...

  16. 46 CFR 116.520 - Emergency evacuation plan.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Emergency evacuation plan. 116.520 Section 116.520... ARRANGEMENT Escape and Embarkation Station Requirements § 116.520 Emergency evacuation plan. The owner or managing operator shall prepare an evacuation plan that must: (a) Identify possible casualties involving...

  17. 46 CFR 116.520 - Emergency evacuation plan.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Emergency evacuation plan. 116.520 Section 116.520... ARRANGEMENT Escape and Embarkation Station Requirements § 116.520 Emergency evacuation plan. The owner or managing operator shall prepare an evacuation plan that must: (a) Identify possible casualties involving...

  18. 46 CFR 116.520 - Emergency evacuation plan.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Emergency evacuation plan. 116.520 Section 116.520... ARRANGEMENT Escape and Embarkation Station Requirements § 116.520 Emergency evacuation plan. The owner or managing operator shall prepare an evacuation plan that must: (a) Identify possible casualties involving...

  19. 42 CFR 136.116 - Reports.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 42 Public Health 1 2012-10-01 2012-10-01 false Reports. 136.116 Section 136.116 Public Health... and Services § 136.116 Reports. In addition to the reporting and information requirements provided in... Federal financial assistance shall make such reports and information available to the Indian people served...

  20. 42 CFR 136.116 - Reports.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Reports. 136.116 Section 136.116 Public Health... and Services § 136.116 Reports. In addition to the reporting and information requirements provided in... Federal financial assistance shall make such reports and information available to the Indian people served...

  1. 42 CFR 136.116 - Reports.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 42 Public Health 1 2011-10-01 2011-10-01 false Reports. 136.116 Section 136.116 Public Health... and Services § 136.116 Reports. In addition to the reporting and information requirements provided in... Federal financial assistance shall make such reports and information available to the Indian people served...

  2. 28 CFR 0.116 - Board of Immigration Appeals.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Board of Immigration Appeals. 0.116 Section 0.116 Judicial Administration DEPARTMENT OF JUSTICE ORGANIZATION OF THE DEPARTMENT OF JUSTICE Executive Office for Immigration Review § 0.116 Board of Immigration Appeals. The Board of Immigration...

  3. Cloning of an SNF2/SWI2-related protein that binds specifically to the SPH motifs of the SV40 enhancer and to the HIV-1 promoter.

    PubMed

    Sheridan, P L; Schorpp, M; Voz, M L; Jones, K A

    1995-03-03

    We have isolated a human cDNA clone encoding HIP116, a protein that binds to the SPH repeats of the SV40 enhancer and to the TATA/inhibitor region of the human immunodeficiency virus (HIV)-1 promoter. The predicted HIP116 protein is related to the yeast SNF2/SWI2 transcription factor and to other members of this extended family and contains seven domains similar to those found in the vaccinia NTP1 ATPase. Interestingly, HIP116 also contains a C3HC4 zinc-binding motif (RING finger) interspersed between the ATPase motifs in an arrangement similar to that found in the yeast RAD5 and RAD16 proteins. The HIP116 amino terminus is unique among the members of this family, and houses a specific DNA-binding domain. Antiserum raised against HIP116 recognizes a 116-kDa nuclear protein in Western blots and specifically supershifts SV40 and HIV-1 protein-DNA complexes in gel shift experiments. The binding site for HIP116 on the SV40 enhancer directly overlaps the site for TEF-1, and like TEF-1, binding of HIP116 to the SV40 enhancer is destroyed by mutations that inhibit SPH enhancer activity in vivo. Purified fractions of HIP116 display strong ATPase activity that is preferentially stimulated by SPH DNA and can be inhibited specifically by antibodies to HIP116. These findings suggest that HIP116 might affect transcription, directly or indirectly, by acting as a DNA binding site-specific ATPase.

  4. 34 CFR 668.116 - Hearing.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 34 Education 3 2011-07-01 2011-07-01 false Hearing. 668.116 Section 668.116 Education Regulations... Program Review Determinations § 668.116 Hearing. (a) A hearing is a process conducted by the hearing official whereby an orderly presentation of arguments and evidence is made by the parties. (b) The hearing...

  5. 45 CFR 46.116 - General requirements for informed consent.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 45 Public Welfare 1 2012-10-01 2012-10-01 false General requirements for informed consent. 46.116 Section 46.116 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION PROTECTION OF HUMAN SUBJECTS Basic HHS Policy for Protection of Human Research Subjects § 46.116 General requirements...

  6. 45 CFR 46.116 - General requirements for informed consent.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 45 Public Welfare 1 2014-10-01 2014-10-01 false General requirements for informed consent. 46.116 Section 46.116 Public Welfare Department of Health and Human Services GENERAL ADMINISTRATION PROTECTION OF HUMAN SUBJECTS Basic HHS Policy for Protection of Human Research Subjects § 46.116 General requirements...

  7. 45 CFR 46.116 - General requirements for informed consent.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 45 Public Welfare 1 2013-10-01 2013-10-01 false General requirements for informed consent. 46.116 Section 46.116 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION PROTECTION OF HUMAN SUBJECTS Basic HHS Policy for Protection of Human Research Subjects § 46.116 General requirements...

  8. 29 CFR 102.116 - Signature of orders.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 2 2010-07-01 2010-07-01 false Signature of orders. 102.116 Section 102.116 Labor Regulations Relating to Labor NATIONAL LABOR RELATIONS BOARD RULES AND REGULATIONS, SERIES 8 Certification and Signature of Documents § 102.116 Signature of orders. The executive secretary or the associate executive...

  9. 40 CFR 265.116 - Survey plat.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 26 2014-07-01 2014-07-01 false Survey plat. 265.116 Section 265.116... and Post-Closure § 265.116 Survey plat. No later than the submission of the certification of closure..., or the authority with jurisdiction over local land use, and to the Regional Administrator, a survey...

  10. 40 CFR 264.116 - Survey plat.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 26 2011-07-01 2011-07-01 false Survey plat. 264.116 Section 264.116...-Closure § 264.116 Survey plat. No later than the submission of the certification of closure of each... authority with jurisdiction over local land use, and to the Regional Administrator, a survey plat indicating...

  11. 40 CFR 264.116 - Survey plat.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Survey plat. 264.116 Section 264.116...-Closure § 264.116 Survey plat. No later than the submission of the certification of closure of each... authority with jurisdiction over local land use, and to the Regional Administrator, a survey plat indicating...

  12. 40 CFR 265.116 - Survey plat.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 27 2012-07-01 2012-07-01 false Survey plat. 265.116 Section 265.116... and Post-Closure § 265.116 Survey plat. No later than the submission of the certification of closure..., or the authority with jurisdiction over local land use, and to the Regional Administrator, a survey...

  13. 40 CFR 264.116 - Survey plat.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 26 2014-07-01 2014-07-01 false Survey plat. 264.116 Section 264.116...-Closure § 264.116 Survey plat. No later than the submission of the certification of closure of each... authority with jurisdiction over local land use, and to the Regional Administrator, a survey plat indicating...

  14. 40 CFR 265.116 - Survey plat.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 26 2011-07-01 2011-07-01 false Survey plat. 265.116 Section 265.116... and Post-Closure § 265.116 Survey plat. No later than the submission of the certification of closure..., or the authority with jurisdiction over local land use, and to the Regional Administrator, a survey...

  15. 40 CFR 264.116 - Survey plat.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 27 2012-07-01 2012-07-01 false Survey plat. 264.116 Section 264.116...-Closure § 264.116 Survey plat. No later than the submission of the certification of closure of each... authority with jurisdiction over local land use, and to the Regional Administrator, a survey plat indicating...

  16. 40 CFR 265.116 - Survey plat.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 27 2013-07-01 2013-07-01 false Survey plat. 265.116 Section 265.116... and Post-Closure § 265.116 Survey plat. No later than the submission of the certification of closure..., or the authority with jurisdiction over local land use, and to the Regional Administrator, a survey...

  17. 40 CFR 265.116 - Survey plat.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Survey plat. 265.116 Section 265.116... and Post-Closure § 265.116 Survey plat. No later than the submission of the certification of closure..., or the authority with jurisdiction over local land use, and to the Regional Administrator, a survey...

  18. 40 CFR 264.116 - Survey plat.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 27 2013-07-01 2013-07-01 false Survey plat. 264.116 Section 264.116...-Closure § 264.116 Survey plat. No later than the submission of the certification of closure of each... authority with jurisdiction over local land use, and to the Regional Administrator, a survey plat indicating...

  19. 42 CFR 433.116 - FFP for operation of mechanized claims processing and information retrieval systems.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... and information retrieval systems. 433.116 Section 433.116 Public Health CENTERS FOR MEDICARE... FISCAL ADMINISTRATION Mechanized Claims Processing and Information Retrieval Systems § 433.116 FFP for operation of mechanized claims processing and information retrieval systems. (a) Subject to paragraph (j) of...

  20. 42 CFR 433.116 - FFP for operation of mechanized claims processing and information retrieval systems.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... and information retrieval systems. 433.116 Section 433.116 Public Health CENTERS FOR MEDICARE... FISCAL ADMINISTRATION Mechanized Claims Processing and Information Retrieval Systems § 433.116 FFP for operation of mechanized claims processing and information retrieval systems. (a) Subject to paragraph (j) of...

  1. 42 CFR 433.116 - FFP for operation of mechanized claims processing and information retrieval systems.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... and information retrieval systems. 433.116 Section 433.116 Public Health CENTERS FOR MEDICARE... FISCAL ADMINISTRATION Mechanized Claims Processing and Information Retrieval Systems § 433.116 FFP for operation of mechanized claims processing and information retrieval systems. (a) Subject to paragraph (j) of...

  2. 42 CFR 433.116 - FFP for operation of mechanized claims processing and information retrieval systems.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... and information retrieval systems. 433.116 Section 433.116 Public Health CENTERS FOR MEDICARE... FISCAL ADMINISTRATION Mechanized Claims Processing and Information Retrieval Systems § 433.116 FFP for operation of mechanized claims processing and information retrieval systems. (a) Subject to paragraph (j) of...

  3. 46 CFR 116.970 - Protection against hot piping.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Protection against hot piping. 116.970 Section 116.970 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE... ARRANGEMENT Rails and Guards § 116.970 Protection against hot piping. Piping, including valves, pipe fittings...

  4. 46 CFR 116.970 - Protection against hot piping.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Protection against hot piping. 116.970 Section 116.970 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE... ARRANGEMENT Rails and Guards § 116.970 Protection against hot piping. Piping, including valves, pipe fittings...

  5. 46 CFR 116.970 - Protection against hot piping.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Protection against hot piping. 116.970 Section 116.970 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE... ARRANGEMENT Rails and Guards § 116.970 Protection against hot piping. Piping, including valves, pipe fittings...

  6. 46 CFR 116.970 - Protection against hot piping.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Protection against hot piping. 116.970 Section 116.970 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE... ARRANGEMENT Rails and Guards § 116.970 Protection against hot piping. Piping, including valves, pipe fittings...

  7. 46 CFR 116.970 - Protection against hot piping.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Protection against hot piping. 116.970 Section 116.970 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE... ARRANGEMENT Rails and Guards § 116.970 Protection against hot piping. Piping, including valves, pipe fittings...

  8. 42 CFR 435.116 - Pregnant women.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 42 Public Health 4 2013-10-01 2013-10-01 false Pregnant women. 435.116 Section 435.116 Public..., AND AMERICAN SAMOA Mandatory Coverage Mandatory Coverage of Pregnant Women, Children Under 19, and Newborn Children § 435.116 Pregnant women. (a) Basis. This section implements sections 1902(a)(10)(A)(i...

  9. 42 CFR 435.116 - Pregnant women.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 42 Public Health 4 2014-10-01 2014-10-01 false Pregnant women. 435.116 Section 435.116 Public..., AND AMERICAN SAMOA Mandatory Coverage Mandatory Coverage of Pregnant Women, Children Under 19, and Newborn Children § 435.116 Pregnant women. (a) Basis. This section implements sections 1902(a)(10)(A)(i...

  10. 21 CFR 133.116 - Low sodium cheddar cheese.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Low sodium cheddar cheese. 133.116 Section 133.116... FOR HUMAN CONSUMPTION CHEESES AND RELATED CHEESE PRODUCTS Requirements for Specific Standardized Cheese and Related Products § 133.116 Low sodium cheddar cheese. Low sodium cheddar cheese is the food...

  11. 21 CFR 133.116 - Low sodium cheddar cheese.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Low sodium cheddar cheese. 133.116 Section 133.116... FOR HUMAN CONSUMPTION CHEESES AND RELATED CHEESE PRODUCTS Requirements for Specific Standardized Cheese and Related Products § 133.116 Low sodium cheddar cheese. Low sodium cheddar cheese is the food...

  12. 40 CFR 116.3 - Definitions.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 23 2013-07-01 2013-07-01 false Definitions. 116.3 Section 116.3 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS DESIGNATION OF HAZARDOUS SUBSTANCES § 116.3 Definitions. As used in this part, all terms shall have the meaning defined in the Act and...

  13. 40 CFR 116.3 - Definitions.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Definitions. 116.3 Section 116.3 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS DESIGNATION OF HAZARDOUS SUBSTANCES § 116.3 Definitions. As used in this part, all terms shall have the meaning defined in the Act and...

  14. 23 CFR 636.116 - What organizational conflict of interest requirements apply to design-build projects?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... apply to design-build projects? 636.116 Section 636.116 Highways FEDERAL HIGHWAY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION ENGINEERING AND TRAFFIC OPERATIONS DESIGN-BUILD CONTRACTING General § 636.116 What organizational conflict of interest requirements apply to design-build projects? (a) State...

  15. 23 CFR 636.116 - What organizational conflict of interest requirements apply to design-build projects?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... apply to design-build projects? 636.116 Section 636.116 Highways FEDERAL HIGHWAY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION ENGINEERING AND TRAFFIC OPERATIONS DESIGN-BUILD CONTRACTING General § 636.116 What organizational conflict of interest requirements apply to design-build projects? (a) State...

  16. 23 CFR 636.116 - What organizational conflict of interest requirements apply to design-build projects?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... apply to design-build projects? 636.116 Section 636.116 Highways FEDERAL HIGHWAY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION ENGINEERING AND TRAFFIC OPERATIONS DESIGN-BUILD CONTRACTING General § 636.116 What organizational conflict of interest requirements apply to design-build projects? (a) State...

  17. 23 CFR 636.116 - What organizational conflict of interest requirements apply to design-build projects?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... apply to design-build projects? 636.116 Section 636.116 Highways FEDERAL HIGHWAY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION ENGINEERING AND TRAFFIC OPERATIONS DESIGN-BUILD CONTRACTING General § 636.116 What organizational conflict of interest requirements apply to design-build projects? (a) State...

  18. 9 CFR 116.7 - Test records.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Test records. 116.7 Section 116.7..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS RECORDS AND REPORTS § 116.7 Test records. Detailed records of all tests conducted on each serial and each subserial shall be maintained by the...

  19. 9 CFR 116.7 - Test records.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Test records. 116.7 Section 116.7..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS RECORDS AND REPORTS § 116.7 Test records. Detailed records of all tests conducted on each serial and each subserial shall be maintained by the...

  20. 20 CFR 632.116 - Conflict of interest.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Conflict of interest. 632.116 Section 632.116... EMPLOYMENT AND TRAINING PROGRAMS Prevention of Fraud and Program Abuse § 632.116 Conflict of interest. (a) No... contractor shall avoid personal and organizational conflict of interest in awarding financial assistance and...

Some links on this page may take you to non-federal websites. Their policies may differ from this site.
Top