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Artículos de revistas sobre el tema "INSPOC"

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Autor: Grafiati

Publicado: 26 de julio de 2024

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1

Hirano, Yoshinobu, Takayuki Nakagomi, Akiko Nakano-Doi, Shuji Kubo, Yusuke Minato, Toshinori Sawano, Masafumi Sakagami y Kenzo Tsuzuki. "Microglia Negatively Regulate the Proliferation and Neuronal Differentiation of Neural Stem/Progenitor Cells Isolated from Poststroke Mouse Brains". Cells 12, n.º 16 (10 de agosto de 2023): 2040. http://dx.doi.org/10.3390/cells12162040.

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We previously demonstrated that neural stem/progenitor cells (NSPCs) were induced within and around the ischemic areas in a mouse model of ischemic stroke. These injury/ischemia-induced NSPCs (iNSPCs) differentiated to electrophysiologically functional neurons in vitro, indicating the presence of a self-repair system following injury. However, during the healing process after stroke, ischemic areas were gradually occupied by inflammatory cells, mainly microglial cells/macrophages (MGs/MΦs), and neurogenesis rarely occurred within and around the ischemic areas. Therefore, to achieve neural regeneration by utilizing endogenous iNSPCs, regulation of MGs/MΦs after an ischemic stroke might be necessary. To test this hypothesis, we used iNSPCs isolated from the ischemic areas after a stroke in our mouse model to investigate the role of MGs/MΦs in iNSPC regulation. In coculture experiments, we show that the presence of MGs/MΦs significantly reduces not only the proliferation but also the differentiation of iNSPCs toward neuronal cells, thereby preventing neurogenesis. These effects, however, are mitigated by MG/MΦ depletion using clodronate encapsulated in liposomes. Additionally, gene ontology analysis reveals that proliferation and neuronal differentiation are negatively regulated in iNSPCs cocultured with MGs/MΦs. These results indicate that MGs/MΦs negatively impact neurogenesis via iNSPCs, suggesting that the regulation of MGs/MΦs is essential to achieve iNSPC-based neural regeneration following an ischemic stroke.

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2

Lakin-Thomas, P. L. "Effects of inositol starvation on the levels of inositol phosphates and inositol lipids in Neurospora crassa". Biochemical Journal 292, n.º 3 (15 de junio de 1993): 805–11. http://dx.doi.org/10.1042/bj2920805.

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An inositol-requiring strain of Neurospora crassa was labelled during growth in liquid medium with [3H]inositol, and the levels of inositol phosphates and phosphoinositides were determined under inositol-sufficient and inositol-starved conditions. Because the mutant has an absolute requirement for inositol, the total mass of inositol-containing compounds could be determined. Inositol-containing lipids were identified by deacylation and co-migration with standards on h.p.l.c.; PtdIns3P, PtdIns4P, and PtdIns(4,5)P2 were found in approximately equal amounts, in addition to large amounts of PtdIns. Inositol starvation decreased the level of PtdIns to 10% of the sufficient level, and decreased the levels of the other phosphoinositides to about 25%. A number of inositol phosphates were found, including several InsP3s, InsP4s and InsP5s and phytic acid. Ins(1,4,5)P3 was identified by co-migration with standards on h.p.l.c. and by digestion with inositol phosphomonoesterase. High concentrations of all inositol phosphates were found in the extracellular medium in inositol-starved cultures. Inositol starvation on both liquid and solid agar media decreased the intracellular levels of some inositol phosphates, but increased the levels of phytic acid and several other inositol phosphates which may be its precursors and/or breakdown products. These results may indicate that inositol starvation induces phytic acid synthesis as a protection against the free-radical production and lipid peroxidation characteristic of inositol-less death.

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3

Tonk, Miray, James J. Valdés, Alejandro Cabezas-Cruz y Andreas Vilcinskas. "Potent Activity of Hybrid Arthropod Antimicrobial Peptides Linked by Glycine Spacers". International Journal of Molecular Sciences 22, n.º 16 (18 de agosto de 2021): 8919. http://dx.doi.org/10.3390/ijms22168919.

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Arthropod antimicrobial peptides (AMPs) offer a promising source of new leads to address the declining number of novel antibiotics and the increasing prevalence of multidrug-resistant bacterial pathogens. AMPs with potent activity against Gram-negative bacteria and distinct modes of action have been identified in insects and scorpions, allowing the discovery of AMP combinations with additive and/or synergistic effects. Here, we tested the synergistic activity of two AMPs, from the dung beetle Copris tripartitus (CopA3) and the scorpion Heterometrus petersii (Hp1090), against two strains of Escherichia coli. We also tested the antibacterial activity of two hybrid peptides generated by joining CopA3 and Hp1090 with linkers comprising two (InSco2) or six (InSco6) glycine residues. We found that CopA3 and Hp1090 acted synergistically against both bacterial strains, and the hybrid peptide InSco2 showed more potent bactericidal activity than the parental AMPs or InSco6. Molecular dynamics simulations revealed that the short linker stabilizes an N-terminal 310-helix in the hybrid peptide InSco2. This secondary structure forms from a coil region that interacts with phosphatidylethanolamine in the membrane bilayer model. The highest concentration of the hybrid peptides used in this study was associated with stronger hemolytic activity than equivalent concentrations of the parental AMPs. As observed for CopA3, the increasing concentration of InSco2 was also cytotoxic to BHK-21 cells. We conclude that AMP hybrids linked by glycine spacers display potent antibacterial activity and that the cytotoxic activity can be modulated by adjusting the nature of the linker peptide, thus offering a strategy to produce hybrid peptides as safe replacements or adjuncts for conventional antibiotic therapy.

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4

Nishiyama, Ryo, Takayuki Nakagomi, Akiko Nakano-Doi, Yoji Kuramoto, Masahiro Tsuji y Shinichi Yoshimura. "Neonatal Brains Exhibit Higher Neural Reparative Activities than Adult Brains in a Mouse Model of Ischemic Stroke". Cells 13, n.º 6 (15 de marzo de 2024): 519. http://dx.doi.org/10.3390/cells13060519.

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The neonatal brain is substantially more resistant to various forms of injury than the mature brain. For instance, the prognosis following ischemic stroke is generally poor in the elderly but favorable in neonates. Identifying the cellular and molecular mechanisms underlying reparative activities in the neonatal brain after ischemic injury may provide feasible targets for therapeutic interventions in adults. To this end, we compared the reparative activities in postnatal day 13 and adult (8–12-week-old) mouse brain following middle cerebral artery occlusion. Immunohistochemistry revealed considerably greater generation of ischemia-induced neural stem/progenitor cells (iNSPCs) expressing nestin or Sox2 in ischemic areas of the neonatal brain. The iNSPCs isolated from the neonatal brain also demonstrated greater proliferative activity than those isolated from adult mice. In addition, genes associated with neuronal differentiation were enriched in iNSPCs isolated from the neonatal brain according to microarray and gene ontogeny analyses. Immunohistochemistry further revealed considerably greater production of newborn doublecortin+ neurons at the sites of ischemic injury in the neonatal brain compared to the adult brain. These findings suggest that greater iNSPC generation and neurogenic differentiation capacities contribute to the superior regeneration of the neonatal brain following ischemia. Together, our findings may help identify therapeutic targets for enhancing the reparative potential of the adult brain following stroke.

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5

Abad, Julián Martín. "Inspice signum". Cultura, Vol. 33 (1 de diciembre de 2014): 45–73. http://dx.doi.org/10.4000/cultura.2354.

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6

Darabseh, Mohammad. "Lean Applications in Construction". U.Porto Journal of Engineering 5, n.º 2 (29 de noviembre de 2019): 29–37. http://dx.doi.org/10.24840/2183-6493_005.002_0004.

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Lean Construction is one of the methods used to improve control over construction projects by eliminating waste in time and materials. Lean Construction is an adaptation from the Lean Manufacturing principles to the construction industry. The purpose of this article was to review the case studies published in 2018 in the Inspec database to find out where Lean is being implemented and how. The article approach by setting the search criteria first and then inspect the result to find the non-related results and eliminate it. After that, the article was reviewed and summarized. The article verifies each article finding; however Lean Construction and Building Information Modeling (BIM) are relatively new for the construction industry and they still need more time to be adopted widely and to be applied on a lower-cost budget. However, the discussed cases show a promising future for these technologies.

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7

Karanjai, Aruna y Usha Mujoo‐Munshi. "The document supply service of INSDOC: an overview". Interlending & Document Supply 17, n.º 2 (febrero de 1989): 49–55. http://dx.doi.org/10.1108/eum0000000003458.

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8

Deaves, J. C. y J. E. Pache. "Chemical and numerical indexing for the INSPEC Database". Indexer 16, n.º 3 (abril de 1989): 163–67. http://dx.doi.org/10.3828/indexer.1989.16.3.6.

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9

Pritchard, R. "Return and inspect". Manufacturing Engineer 80, n.º 6 (1 de diciembre de 2001): 252–54. http://dx.doi.org/10.1049/me:20010606.

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10

Sherwin, D. J. "Inspect or monitor?" Engineering Costs and Production Economics 18, n.º 3 (enero de 1990): 223–31. http://dx.doi.org/10.1016/0167-188x(90)90124-z.

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11

Not applicable. "INSPEC Database To Be Available on FirstSearch and EPIC". Issues in Science and Technology Librarianship, n.º 8 (15 de diciembre de 1993): 356–94. http://dx.doi.org/10.29173/istl1735.

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12

McAllister III, James Thomas y Nancy Stephanie Diaz. "Hosting Inspec on Engineering Village or Web of Science". Reference Services Review 48, n.º 2 (13 de abril de 2020): 307–17. http://dx.doi.org/10.1108/rsr-08-2019-0050.

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Purpose As library budgets continue to constrict, librarians will need to become more familiar with comparing database host platforms. This paper aims to compare Inspec on Elsevier’s Engineering Village (EV) and Clarivate’s Web of Science (WOS) from a novice user experience. The main objectives are to identify some R1 institutions that subscribe to Inspec and highlight some of the key differences between the two platforms. Design/methodology/approach Information on Inspec was gathered from various sources such as the home website, IET, and the host platform websites of Elsevier and Clarivate Analytics. Additional evidence was also collected from brochures and guides to help illustrate some of the main features and differences that novice users would be familiar with. Findings Most institutions subscribe to Inspec via the EV platform. Results from the study conclude that EV was the choice over WOS for hosting Inspec owing to a more user-friendly interface, potential lower cost and faster platform updates in response to meeting user needs. Research limitations/implications As database platforms change over time, feature areas such as content, interface and features remain important for information professionals and librarians to stay current with those changes. Also, this work can help librarians with understanding the planning and developing of a process for comparing identical or similar databases on different platforms. Originality/value Much of the literature focuses on the unfamiliar details and not so much on the novice user. This paper provides a unique perspective in how a novice user would prefer the attributes of one host platform over the other. Additionally, the same review criteria can be applied in other subjects and disciplines.

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13

Bardin, D. Yu, L. V. Kalinovskaya y E. D. Uglov. "Zγγγ→0 Processes inSANC". Ядерная физика 76, n.º 11 (2013): 1404–9. http://dx.doi.org/10.7868/s0044002713110032.

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14

Learner, Sue. "Inspect school nurseries separately?" Practical Pre-School 2014, n.º 160 (mayo de 2014): 4–6. http://dx.doi.org/10.12968/prps.2014.5.160.4a.

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15

Browne, Maureen. "An insight into Inspex". Production Engineer 64, n.º 3 (1985): 32. http://dx.doi.org/10.1049/tpe.1985.0072.

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16

Ikoma, Takashi. "Properties of Gallium Arsenide, Datareviews Series No. 2 (INSPEC, 1986)". MRS Bulletin 13, n.º 3 (marzo de 1988): 48. http://dx.doi.org/10.1557/s0883769400066203.

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17

Dhaka, R. P. S. y Kamlesh Arora. "Privatisation of S&T Information Services : The INSDOC Experience". DESIDOC Bulletin of Information Technology 15, n.º 5 (1 de septiembre de 1995): 7–10. http://dx.doi.org/10.14429/dbit.15.5.3200.

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18

Payne, Matthew Thomas. "War Bytes: The Critique of Militainment inSpec Ops: The Line". Critical Studies in Media Communication 31, n.º 4 (13 de febrero de 2014): 265–82. http://dx.doi.org/10.1080/15295036.2014.881518.

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19

Traynor, Kate. "NABP to inspect compounding pharmacies". American Journal of Health-System Pharmacy 70, n.º 2 (1 de enero de 2013): 88–90. http://dx.doi.org/10.2146/news130006.

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20

Tajiri, Hisao y Mario Dinis-Ribeiro. "How I inspect the stomach". Endoscopy 51, n.º 06 (3 de abril de 2019): 571–73. http://dx.doi.org/10.1055/a-0876-1873.

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21

Kenzie, Bryan. "Robots to inspect welds underwater". Industrial Robot: An International Journal 21, n.º 3 (junio de 1994): 13–14. http://dx.doi.org/10.1108/eum0000000004150.

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22

Hamann, D. R. "Diffusion of Atomic Oxygen inSiO2". Physical Review Letters 81, n.º 16 (19 de octubre de 1998): 3447–50. http://dx.doi.org/10.1103/physrevlett.81.3447.

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23

Afanas’ev, V. V. y A. Stesmans. "Photoionization of silicon particles inSiO2". Physical Review B 59, n.º 3 (15 de enero de 1999): 2025–34. http://dx.doi.org/10.1103/physrevb.59.2025.

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24

Copperman, K., K. Canepa, K. Murphy, M. Acosta-La Greca, L. Chuang y A. B. Copperman. "People respect what you inspect". Fertility and Sterility 92, n.º 3 (septiembre de 2009): S61—S62. http://dx.doi.org/10.1016/j.fertnstert.2009.07.239.

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25

Carson, Deirdre. "Allowing RVNs to inspect practices". Veterinary Record 184, n.º 21 (23 de mayo de 2019): 655.2–655. http://dx.doi.org/10.1136/vr.l3100.

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26

Hamaker, Ellen L. "Commentary: How to Inspect Fruit". Measurement: Interdisciplinary Research & Perspective 5, n.º 4 (4 de diciembre de 2007): 250–53. http://dx.doi.org/10.1080/15366360701775961.

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27

Fullstone, Mark. "Trainable Systems Inspect Food Products". Sensor Review 13, n.º 2 (febrero de 1993): 13–15. http://dx.doi.org/10.1108/eb007900.

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28

McClean, Andrew. "In Focus: Inspecs marches on". Optician 2020, n.º 11 (noviembre de 2020): 232515–1. http://dx.doi.org/10.12968/opti.2020.11.232515.

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29

Tajiri, Hisao y Mário Dinis-Ribeiro. "How I inspect the stomach". Gastrointestinal Endoscopy 89, n.º 6 (junio de 2019): 1106–8. http://dx.doi.org/10.1016/j.gie.2019.02.003.

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30

Suvanov, Sardor. "SOME ISSUES OF HUMAN RIGHTS IN THE UNIVERSAL DECLARATION OF HUMAN RIGHTS". Jurisprudence 1, n.º 6 (15 de diciembre de 2021): 209–15. http://dx.doi.org/10.51788/tsul.jurisprudence.1.6./pfmj9556.

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This article analyzes norms aimed at protecting human rights set out in the Universal Declaration of Human Rights. The civil and personal rights enshrined in the Declaration, as well as economic, social and cultural rights, were discussed in detail on the basis of the Constitution of Uzbekistan. In particular, the right to life, the right to liberty and security of person, the prohibition of torture, cruel, inhuman or degrading treatment or punishment, the right to housing, the right to freedom of thought, speech and religion were analyzed. At the same time, this article examines the ongoing reforms in the field of human rights and the existing problems in the protection of human rights. Appropriate proposals and recommendations have been developed to address the problems. In particular, proposals are being put forward regarding strengthening cooperation between government agencies and civil society institutions in the field of human rights, ensuring parliamentary and public control over the implementation of relevant laws, strengthening accountability for human rights violations, expanding the system of human rights training for government officials directly responsible for human rights and to setting up a system of training them.

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31

Levine, Deb, Andrew J. Woodruff, A. Rain Mocello, Jaime Lebrija y Jeffrey D. Klausner. "inSPOT: The First Online STD Partner Notification System Using Electronic Postcards". PLoS Medicine 5, n.º 10 (21 de octubre de 2008): e213. http://dx.doi.org/10.1371/journal.pmed.0050213.

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32

Dickinson, Sarah. "From Avery to INSPEC: A Collaborative Approach to Instruction Across Disciplines". Art Documentation: Journal of the Art Libraries Society of North America 25, n.º 1 (abril de 2006): 43–45. http://dx.doi.org/10.1086/adx.25.1.27949401.

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33

Shirinova, Ekaterina y Gulbakhor Iskandarova. "HUMAN – AT THE INTERSECTION OF CULTURES". Uzbekistan:Language and culture 5, n.º 2 (26 de febrero de 2023): 83–89. http://dx.doi.org/10.52773/tsuull.uzlc.aphil.2023.2.5/nqug6205.

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Understanding the world, a person understands himself. By be-coming aware of himself, he understands the world and his place in it. He feels harmony with the existence in which he lives. This process is carried out through the personal cultural prism of a person, which is created as a result of the environment in which he was born, grew up and matured. That is, a person looks at the world through his beliefs, moral rules, tradi-tions, value system and native language. These situations are explained in science using the terms cultural picture of the world and linguistic picture of the world.This article provides an explanation of the concept of culture, the interaction of cultures, the processes of inculturation and acculturation. Information is given about the main differences between these processes, what changes occur in cultures as a result of the interaction of cultures, and what factors influence these changes. A person’s entire life is analyzed at the intersection of different cultures.

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34

Madhamshettiwar, Piyush B., Stefan R. Maetschke, Melissa J. Davis, Antonio Reverter y Mark A. Ragan. "INsPeCT: INtegrative Platform for Cancer Transcriptomics". Cancer Informatics 13 (enero de 2014): CIN.S13630. http://dx.doi.org/10.4137/cin.s13630.

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The emergence of transcriptomics, fuelled by high-throughput sequencing technologies, has changed the nature of cancer research and resulted in a massive accumulation of data. Computational analysis, integration, and data visualization are now major bottlenecks in cancer biology and translational research. Although many tools have been brought to bear on these problems, their use remains unnecessarily restricted to computational biologists, as many tools require scripting skills, data infrastructure, and powerful computational facilities. New user-friendly, integrative, and automated analytical approaches are required to make computational methods more generally useful to the research community. Here we present INsPeCT (INtegrative Platform for Cancer Transcriptomics), which allows users with basic computer skills to perform comprehensive in-silico analyses of microarray, ChlPseq, and RNA-seq data. INsPeCT supports the selection of interesting genes for advanced functional analysis. Included in its automated workflows are (i) a novel analytical framework, RMaNI (regulatory module network inference), which supports the inference of cancer subtype-specific transcriptional module networks and the analysis of modules; and (ii) WGCNA (weighted gene co-expression network analysis), which infers modules of highly correlated genes across microarray samples, associated with sample traits, eg survival time. INsPeCT is available free of cost from Bioinformatics Resource Australia-EMBL and can be accessed at http://inspect.braembl.org.au .

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35

Hwang, Juck-Joon. "How to Inspect the Dead Body?" Journal of the Korean Medical Association 41, n.º 3 (1998): 240. http://dx.doi.org/10.5124/jkma.1998.41.3.240.

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36

Tuttle, Blair. "Energetics and diffusion of hydrogen inSiO2". Physical Review B 61, n.º 7 (15 de febrero de 2000): 4417–20. http://dx.doi.org/10.1103/physrevb.61.4417.

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37

Vais, Horia, J. Kevin Foskett y Don-On Daniel Mak. "InsP3R channel gating altered by clustering?" Nature 478, n.º 7368 (12 de octubre de 2011): E1—E2. http://dx.doi.org/10.1038/nature10493.

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38

McFeely, F. R., E. Cartier, J. A. Yarmoff y S. A. Joyce. "Low-energy-electron escape lengths inSiO2". Physical Review B 42, n.º 8 (15 de septiembre de 1990): 5191–200. http://dx.doi.org/10.1103/physrevb.42.5191.

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Laermans, C. y V. Keppens. "Origin of the tunneling states inSiO2". Physical Review B 51, n.º 13 (1 de abril de 1995): 8158–63. http://dx.doi.org/10.1103/physrevb.51.8158.

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40

Glasbrenner, K. "Field staff: inspect, initiate enforcement, investigate". JAMA: The Journal of the American Medical Association 254, n.º 16 (25 de octubre de 1985): 2199. http://dx.doi.org/10.1001/jama.254.16.2199.

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41

Hannah, Craig. "Physically Inspect Revenue Water Meters Annually". Opflow 45, n.º 5 (mayo de 2019): 6–7. http://dx.doi.org/10.1002/opfl.1184.

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42

Glasbrenner, Kimberly. "Field staff: inspect, initiate enforcement, investigate". JAMA: The Journal of the American Medical Association 254, n.º 16 (25 de octubre de 1985): 2199. http://dx.doi.org/10.1001/jama.1985.03360160025003.

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43

Ewert, Per. "Inspel från den skandinaviska sekularismens guldfiskskål". Theofilos 14, n.º 13 (28 de febrero de 2022): 1–2. http://dx.doi.org/10.48032/theo/14/1/1.

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Just före publiceringen av denna utgåva av Theofilos har även årets version av världskulturkartan från World Values Survey presenterats. Förändringarna är små jämfört med 2020 års version, men låt mig poängtera några centrala poänger. Det ligger en paradox i att just de protestantiska länderna, med sitt lutherska arv av Skriften och Kristus allena, har genomgått en så stark sekularisering. Där det för några sekler sedan betraktades som närmast otänkbart att föreställa sig tillvaron utan Gud står vi idag istället i ett samhälle där en sekulär livssyn framstår som helt självskriven i offentlig debatt. En granskning av dessa idéförändringar är angelägen att göra för alla instanser i samhället – inte minst för kyrkan.

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44

UCHIBORI, Daisuke, Kazuaki WATANABE, Yosuke SAKURADA y Atsushi ARATAKE. "AUTONOMOUS UAV TO INSPECT COMMUNICATION MANHOLES". Japanese Journal of JSCE 79, n.º 12 (2023): n/a. http://dx.doi.org/10.2208/jscejj.23-00129.

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45

Fujii, M., M. Wada, S. Hayashi y K. Yamamoto. "Infrared absorption inSiO2-Ge composite films: Influences of Ge microcrystals on the longitudinal-optical phonons inSiO2". Physical Review B 46, n.º 24 (15 de diciembre de 1992): 15930–35. http://dx.doi.org/10.1103/physrevb.46.15930.

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46

Correia da Cruz, A. y M. Silva Ribeiro. "RobTank Inspec – in service robotized inspection tool for hazardous products storage tanks". Industrial Robot: An International Journal 32, n.º 2 (abril de 2005): 157–62. http://dx.doi.org/10.1108/01439910510582282.

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47

Zhou, Li Ying, Wen Ge Zuo, Qing Feng Wei y Chang Shou Luo. "Research Profiling of Carbon Nanotubes from 2000 to 2013". Advanced Materials Research 941-944 (junio de 2014): 385–90. http://dx.doi.org/10.4028/www.scientific.net/amr.941-944.385.

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In order to profile the research status of carbon nanotubes (CNTs), and gasp the development direction of the subject, based on the database of INSPEC, with the help of INSPEC thesaurus, using co-word clustering, 8 important subfields of CNTs are identified in this paper. Furthermore, the trends of each subfield with time was showed as well as the journal distribution of each subfield was analyzed. The result should be focused on to provide the institutions, researchers and decision makers of CNTs with useful reference.

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48

Pérez Alvarez-Ossorio, J. R., M. L. Gómez Caridad, M. J. Martín Sempere, C. Galbán Ferrús, M. C. Urdin Caminos y A. L. Sobrado Presa. "La producción de la Universidad española en Física, reflejada en las publicaciones españolas y extranjeras". Revista española de Documentación Científica 14, n.º 4 (16 de julio de 2020): 428. http://dx.doi.org/10.3989/redc.1991.v14.i4.428.

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Se analiza la producción científica de la Universidad española en el campo de la Física, y en el período 1986-88, a través de dos bases de datos: INSPEC, que recoge lo publicado en revistas extranjeras y también, aunque de modo selectivo, en cierto número de revistas españolas; e ICYT, que recoge exhaustivamente todos los trabajos originales publicados en revistas españolas. La comparación entre ambas series de datos permite deducir tendencias de publícación en el extranjero. Asimismo se analizan comparativamente los periodos 1982-85 y 1986-88, con los datos obtenidos de INSPEC.

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Kaznacheyeva, Elena, Vitalie D. Lupu y Ilya Bezprozvanny. "Single-Channel Properties of Inositol (1,4,5)-Trisphosphate Receptor Heterologously Expressed in HEK-293 Cells". Journal of General Physiology 111, n.º 6 (1 de junio de 1998): 847–56. http://dx.doi.org/10.1085/jgp.111.6.847.

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The inositol (1,4,5)-trisphosphate receptor (InsP3R) mediates Ca2+ release from intracellular stores in response to generation of second messenger InsP3. InsP3R was biochemically purified and cloned, and functional properties of native InsP3-gated Ca2+ channels were extensively studied. However, further studies of InsP3R are obstructed by the lack of a convenient functional assay of expressed InsP3R activity. To establish a functional assay of recombinant InsP3R activity, transient heterologous expression of neuronal rat InsP3R cDNA (InsP3R-I, SI− SII+ splice variant) in HEK-293 cells was combined with the planar lipid bilayer reconstitution experiments. Recombinant InsP3R retained specific InsP3 binding properties (Kd = 60 nM InsP3) and were specifically recognized by anti–InsP3R-I rabbit polyclonal antibody. Density of expressed InsP3R-I was at least 20-fold above endogenous InsP3R background and only 2–3-fold lower than InsP3R density in rat cerebellar microsomes. When incorporated into planar lipid bilayers, the recombinant InsP3R formed a functional InsP3-gated Ca2+ channel with 80 pS conductance using 50 mM Ba2+ as a current carrier. Mean open time of recombinant InsP3-gated channels was 3.0 ms; closed dwell time distribution was double exponential and characterized by short (18 ms) and long (130 ms) time constants. Overall, gating and conductance properties of recombinant neuronal rat InsP3R-I were very similar to properties of native rat cerebellar InsP3R recorded in identical experimental conditions. Recombinant InsP3R also retained bell-shaped dependence on cytosolic Ca2+ concentration and allosteric modulation by ATP, similar to native cerebellar InsP3R. The following conclusions are drawn from these results. (a) Rat neuronal InsP3R-I cDNA encodes a protein that is either sufficient to produce InsP3-gated channel with functional properties identical to the properties of native rat cerebellar InsP3R, or it is able to form a functional InsP3-gated channel by forming a complex with proteins endogenously expressed in HEK-293 cells. (b) Successful functional expression of InsP3R in a heterologous expression system provides an opportunity for future detailed structure–function characterization of this vital protein.

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Salisbury, Lutishoor. "Inspec on Two Platforms: Elsevier’s Engineering Village and Clarivate Analytics’ Web of Science". Charleston Advisor 20, n.º 3 (1 de enero de 2019): 5–13. http://dx.doi.org/10.5260/chara.20.3.5.

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