Generic placeholder image

Protein & Peptide Letters

Editor-in-Chief

ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

Mini-Review Article

Database Study on the Expression and Purification of Membrane Proteins

Author(s): Chen-Yan Zhang *, Shi-Qi Zhao , Shi-Long Zhang , Li-Heng Luo, Ding-Chang Liu, Wei-Hang Ding, Dong-Jie Fu, Xu-Dong Deng and Da-Chuan Yin *

Volume 28, Issue 9, 2021

Published on: 15 April, 2021

Page: [972 - 982] Pages: 11

DOI: 10.2174/0929866528666210415120234

Price: $65

Abstract

Membrane proteins are crucial for biological processes, and many of them are important to drug targets. Understanding the three-dimensional structures of membrane proteins are essential to evaluate their bio-function and drug design. High-purity membrane proteins are important for structural determination. Membrane proteins have low yields and are difficult to purify because they tend to aggregate. We summarized membrane protein expression systems, vectors, tags, and detergents, which have deposited in the Protein Data Bank (PDB) in recent four-and-a-half years. Escherichia coli is the most expression system for membrane proteins, and HEK293 cells are the most commonly cell lines for human membrane protein expression. The most frequently vectors are pFastBac1 for alpha-helical membrane proteins, pET28a for beta-barrel membrane proteins, and pTRC99a for monotopic membrane proteins. The most used tag for membrane proteins is the 6×His-tag. FLAG commonly used for alpha-helical membrane proteins, Strep and GST for beta- barrel and monotopic membrane proteins, respectively. The detergents and their concentrations used for alpha-helical, beta-barrel, and monotopic membrane proteins are different, and DDM is commonly used for membrane protein purification. It can guide the expression and purification of membrane proteins, thus contributing to their structure and bio function studying.

Keywords: Membrane protein, expression, purification, database, tags, detergent.

Graphical Abstract

[1]
Lacapère, J.J.; Pebay-Peyroula, E.; Neumann, J.M.; Etchebest, C. Determining membrane protein structures: still a challenge! Trends Biochem. Sci., 2007, 32(6), 259-270.[http://dx.doi.org/10.1016/j.tibs.2007.04.001] [PMID: 17481903]
[2]
Wise, A.; Gearing, K.; Rees, S. Target validation of G-protein coupled receptors. Drug Discov. Today, 2002, 7(4), 235-246.[http://dx.doi.org/10.1016/S1359-6446(01)02131-6] [PMID: 11839521]
[3]
Chayen, N.E.; Saridakis, E. Protein crystallization: from purified protein to diffraction-quality crystal. Nat. Methods, 2008, 5(2), 147-153.[http://dx.doi.org/10.1038/nmeth.f.203] [PMID: 18235435]
[4]
McPherson, A. Protein crystallization. Methods Mol. Biol., 2017, 1607, 17-50.[http://dx.doi.org/10.1007/978-1-4939-7000-1_2] [PMID: 28573568]
[5]
Cuéllar-Cruz, M.; Schneider, D.K.; Stojanoff, V.; Islas, S.R.; Sanchez-Puig, N.; Arreguin-Espinosa, R.; Delgado, J.M.; Moreno, A. Formation of crystalline silica-carbonate biomorphs of alkaline earth metals (Ca, Ba, Sr) from ambient to low temperatures: chemical implications during the primitive earth’s life. Cryst. Growth Des., 2020, 20, 1186-1195.[http://dx.doi.org/10.1021/acs.cgd.9b01473]
[6]
Botha, S.; Baitan, D.; Jungnickel, K.E.J.; Oberthür, D.; Schmidt, C.; Stern, S.; Wiedorn, M.O.; Perbandt, M.; Chapman, H.N.; Betzel, C. De novo protein structure determination by heavy-atom soaking in lipidic cubic phase and SIRAS phasing using serial synchrotron crystallography. IUCrJ, 2018, 5(Pt 5), 524-530.[http://dx.doi.org/10.1107/S2052252518009223] [PMID: 30224955]
[7]
Seddon, A.M.; Curnow, P.; Booth, P.J. Membrane proteins, lipids and detergents: not just a soap opera. Biochim. Biophys. Acta, 2004, 1666(1-2), 105-117.[http://dx.doi.org/10.1016/j.bbamem.2004.04.011] [PMID: 15519311]
[8]
Zhou, Z.R.; Huang, W.; Liu, K.J.; Lin, F.L.; Wang, X.L.; Wang, F.; Jiang, R. Soluble expression, one-step purification and characterization of recombinant human growth hormone fused with ompA3 in Escherichia coli. Protein Pept. Lett., 2021, 28(5), 533-542.[http://dx.doi.org/10.2174/0929866527666201110123426] [PMID: 33172365]
[9]
Braiman, M.S.; Stern, L.J.; Chao, B.H.; Khorana, H.G. Structure-function studies on bacteriorhodopsin. IV. Purification and renaturation of bacterio-opsin polypeptide expressed in Escherichia coli. J. Biol. Chem., 1987, 262(19), 9271-9276.[http://dx.doi.org/10.1016/S0021-9258(18)48076-3] [PMID: 3298254]
[10]
Fukuda, M.; Eshdat, Y.; Tarone, G.; Marchesi, V.T. Isolation and characterization of peptides derived from the cytoplasmic segment of band 3, the predominant intrinsic membrane protein of the human erythrocyte. J. Biol. Chem., 1978, 253(7), 2419-2428.[http://dx.doi.org/10.1016/S0021-9258(17)38090-0] [PMID: 632277]
[11]
Carpenter, E.P.; Beis, K.; Cameron, A.D.; Iwata, S. Overcoming the challenges of membrane protein crystallography. Curr. Opin. Struct. Biol., 2008, 18(5), 581-586.[http://dx.doi.org/10.1016/j.sbi.2008.07.001] [PMID: 18674618]
[12]
Lyons, J.A.; Shahsavar, A.; Paulsen, P.A.; Pedersen, B.P.; Nissen, P. Expression strategies for structural studies of eukaryotic membrane proteins. Curr. Opin. Struct. Biol., 2016, 38, 137-144.[http://dx.doi.org/10.1016/j.sbi.2016.06.011] [PMID: 27362979]
[13]
Rosano, G.L.; Morales, E.S.; Ceccarelli, E.A. New tools for recombinant protein production in Escherichia coli: A 5-year update. Protein Sci., 2019, 28(8), 1412-1422.[http://dx.doi.org/10.1002/pro.3668] [PMID: 31219641]
[14]
Ahmad, M.; Hirz, M.; Pichler, H.; Schwab, H. Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production. Appl. Microbiol. Biotechnol., 2014, 98(12), 5301-5317.[http://dx.doi.org/10.1007/s00253-014-5732-5] [PMID: 24743983]
[15]
Yang, Z.; Zhang, Z. Engineering strategies for enhanced production of protein and bio-products in Pichia pastoris: a review. Biotechnol. Adv., 2018, 36(1), 182-195.[http://dx.doi.org/10.1016/j.biotechadv.2017.11.002] [PMID: 29129652]
[16]
Gottesman, M.M. Chinese hamster ovary cells. Methods Enzymol., 1987, 151, 3-8.[http://dx.doi.org/10.1016/S0076-6879(87)51004-7] [PMID: 3431446]
[17]
Arena, T.A.; Chou, B.; Harms, P.D.; Wong, A.W. An anti-apoptotic HEK293 cell line provides a robust and high titer platform for transient protein expression in bioreactors. MAbs, 2019, 11(5), 977-986.[http://dx.doi.org/10.1080/19420862.2019.1598230] [PMID: 30907238]
[18]
Brödel, A.K.; Sonnabend, A.; Kubick, S. Cell-free protein expression based on extracts from CHO cells. Biotechnol. Bioeng., 2014, 111(1), 25-36.[http://dx.doi.org/10.1002/bit.25013] [PMID: 24018795]
[19]
Gagoski, D.; Polinkovsky, M.E.; Mureev, S.; Kunert, A.; Johnston, W.; Gambin, Y.; Alexandrov, K. Performance benchmarking of four cell-free protein expression systems. Biotechnol. Bioeng., 2016, 113(2), 292-300.[http://dx.doi.org/10.1002/bit.25814] [PMID: 26301602]
[20]
Studier, F.W.; Moffatt, B.A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J. Mol. Biol., 1986, 189(1), 113-130.[http://dx.doi.org/10.1016/0022-2836(86)90385-2] [PMID: 3537305]
[21]
Rosenberg, A.H.; Lade, B.N.; Chui, D.S.; Lin, S.W.; Dunn, J.J.; Studier, F.W. Vectors for selective expression of cloned DNAs by T7 RNA polymerase. Gene, 1987, 56(1), 125-135.[http://dx.doi.org/10.1016/0378-1119(87)90165-X] [PMID: 3315856]
[22]
Studier, F.W.; Rosenberg, A.H.; Dunn, J.J.; Dubendorff, J.W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol., 1990, 185, 60-89.[http://dx.doi.org/10.1016/0076-6879(90)85008-C] [PMID: 2199796]
[23]
Zhang, X.; Yao, Z.; Duan, Y.; Zhang, X.; Shi, J.; Xu, Z. Investigation of specific interactions between T7 promoter and T7 RNA polymerase by force spectroscopy using atomic force microscope. Biochem. J., 2018, 475(1), 319-328.[http://dx.doi.org/10.1042/BCJ20170616] [PMID: 29187520]
[24]
Wang, W.; Li, Y.; Wang, Y.; Shi, C.; Li, C.; Li, Q.; Linhardt, R.J. Bacteriophage T7 transcription system: an enabling tool in synthetic biology. Biotechnol. Adv., 2018, 36(8), 2129-2137.[http://dx.doi.org/10.1016/j.biotechadv.2018.10.001] [PMID: 30290194]
[25]
Shang, H.; Garretson, T.A.; Kumar, C.M.S.; Dieter, R.F.; Cheng, X.W. Improved pFastBac™ donor plasmid vectors for higher protein production using the Bac-to-Bac® baculovirus expression vector system. J. Biotechnol., 2017, 255, 37-46.[http://dx.doi.org/10.1016/j.jbiotec.2017.06.397] [PMID: 28645582]
[26]
Goehring, A.; Lee, C.H.; Wang, K.H.; Michel, J.C.; Claxton, D.P.; Baconguis, I.; Althoff, T.; Fischer, S.; Garcia, K.C.; Gouaux, E. Screening and large-scale expression of membrane proteins in mammalian cells for structural studies. Nat. Protoc., 2014, 9(11), 2574-2585.[http://dx.doi.org/10.1038/nprot.2014.173] [PMID: 25299155]
[27]
Hochuli, E.; Döbeli, H.; Schacher, A. New metal chelate adsorbent selective for proteins and peptides containing neighbouring histidine residues. J. Chromatogr. A, 1987, 411, 177-184.[http://dx.doi.org/10.1016/S0021-9673(00)93969-4] [PMID: 3443622]
[28]
Abdullah, N.; Chase, H.A. Removal of poly-histidine fusion tags from recombinant proteins purified by expanded bed adsorption. Biotechnol. Bioeng., 2005, 92(4), 501-513.[http://dx.doi.org/10.1002/bit.20633] [PMID: 16080185]
[29]
Tovar, C.N.; Odunuga, O.O. Size of Protein is a major factor that affects retention on preparative IMAC columns. Protein J., 2019, 38(1), 76-82.[http://dx.doi.org/10.1007/s10930-018-9803-9] [PMID: 30448899]
[30]
Kutyshenko, V.P.; Mikoulinskaia, G.V.; Chernyshov, S.V.; Yegorov, A.Y.; Prokhorov, D.A.; Uversky, V.N. Effect of C-terminal His-tag and purification routine on the activity and structure of the metalloenzyme, l-alanyl-d-glutamate peptidase of the bacteriophage T5. Int. J. Biol. Macromol., 2019, 124, 810-818.[http://dx.doi.org/10.1016/j.ijbiomac.2018.11.219] [PMID: 30500497]
[31]
Sayari, A.; Mosbah, H.; Verger, R.; Gargouri, Y. The N-terminal His-tag affects the enantioselectivity of staphylococcal lipases: a monolayer study. J. Colloid Interface Sci., 2007, 313(1), 261-267.[http://dx.doi.org/10.1016/j.jcis.2007.04.053] [PMID: 17532333]
[32]
Terpe, K. Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. Appl. Microbiol. Biotechnol., 2003, 60(5), 523-533.[http://dx.doi.org/10.1007/s00253-002-1158-6] [PMID: 12536251]
[33]
Schmidt, T.G.M.; Koepke, J.; Frank, R.; Skerra, A. Molecular interaction between the Strep-tag affinity peptide and its cognate target, streptavidin. J. Mol. Biol., 1996, 255(5), 753-766.[http://dx.doi.org/10.1006/jmbi.1996.0061] [PMID: 8636976]
[34]
Schmidt, T.G.M.; Skerra, A. The Strep-tag system for one-step purification and high-affinity detection or capturing of proteins. Nat. Protoc., 2007, 2(6), 1528-1535.[http://dx.doi.org/10.1038/nprot.2007.209] [PMID: 17571060]
[35]
Hopp, T.P.; Prickett, K.S.; Price, V.L.; Libby, R.T.; March, C.J.; Cerretti, D.P.; Urdal, D.L.; Conlon, P.J. A short polypeptide marker sequence useful for recombinant protein identification and purification. Bio-Biotechnol., 1988, 6, 1204-1210.[http://dx.doi.org/10.1038/nbt1088-1204]
[36]
Smith, D.B.; Johnson, K.S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene, 1988, 67(1), 31-40.[http://dx.doi.org/10.1016/0378-1119(88)90005-4] [PMID: 3047011]
[37]
Wang, X.; Guo, T.; Chen, J.; Li, X.; Zhou, Y.; Pan, Z. Covalent and selective immobilization of GST fusion proteins with fluorophosphonate-based probes. Chem. Commun. (Camb.), 2018, 54(37), 4661-4664.[http://dx.doi.org/10.1039/C7CC08888D] [PMID: 29542741]
[38]
Wolfe, A.J.; Gugel, J.F.; Chen, M.; Movileanu, L. Detergent desorption of membrane proteins exhibits two kinetic phases. J. Phys. Chem. Lett., 2018, 9(8), 1913-1919.[http://dx.doi.org/10.1021/acs.jpclett.8b00549] [PMID: 29595981]
[39]
Le Bon, C.; Della Pia, E.A.; Giusti, F.; Lloret, N.; Zoonens, M.; Martinez, K.L.; Popot, J.L. Synthesis of an oligonucleotide-derivatized amphipol and its use to trap and immobilize membrane proteins. Nucleic Acids Res., 2014, 42(10), e83.[http://dx.doi.org/10.1093/nar/gku250] [PMID: 24744236]
[40]
Giusti, F.; Kessler, P.; Hansen, R.W.; Della Pia, E.A.; Le Bon, C.; Mourier, G.; Popot, J.L.; Martinez, K.L.; Zoonens, M. Synthesis of a polyhistidine-bearing amphipol and its use for immobilizing membrane proteins. Biomacromolecules, 2015, 16(12), 3751-3761.[http://dx.doi.org/10.1021/acs.biomac.5b01010] [PMID: 26492302]
[41]
Le Bon, C.; Marconnet, A.; Masscheleyn, S.; Popot, J.L.; Zoonens, M. Folding and stabilizing membrane proteins in amphipol A8-35. Methods, 2018, 147, 95-105.[http://dx.doi.org/10.1016/j.ymeth.2018.04.012] [PMID: 29678587]
[42]
Denisov, I.G.; Sligar, S.G. Nanodiscs for structural and functional studies of membrane proteins. Nat. Struct. Mol. Biol., 2016, 23(6), 481-486.[http://dx.doi.org/10.1038/nsmb.3195] [PMID: 27273631]
[43]
Nollert, P. Membrane protein crystallization in amphiphile phases: practical and theoretical considerations. Prog. Biophys. Mol. Biol., 2005, 88(3), 339-357.[http://dx.doi.org/10.1016/j.pbiomolbio.2004.07.006] [PMID: 15652249]
[44]
Lee, Y.C.; Bååth, J.A.; Bastle, R.M.; Bhattacharjee, S.; Cantoria, M.J.; Dornan, M.; Gamero-Estevez, E.; Ford, L.; Halova, L.; Kernan, J.; Kürten, C.; Li, S.; Martinez, J.; Sachan, N.; Sarr, M.; Shan, X.; Subramanian, N.; Rivera, K.; Pappin, D.; Lin, S.H. Impact of detergents on membrane protein complex isolation. J. Proteome Res., 2018, 17(1), 348-358.[http://dx.doi.org/10.1021/acs.jproteome.7b00599] [PMID: 29110486]
[45]
Opitz, S.; Hannika, F.; Krüger, T.; Rhode, H. The removal of Triton X-100 by dialysis is feasible! Anal. Bioanal. Chem., 2015, 407(4), 1107-1118.[http://dx.doi.org/10.1007/s00216-014-8333-3] [PMID: 25424179]
[46]
le Maire, M.; Arnou, B.; Olesen, C.; Georgin, D.; Ebel, C.; Møller, J.V. Gel chromatography and analytical ultracentrifugation to determine the extent of detergent binding and aggregation, and stokes radius of membrane proteins using sarcoplasmic reticulum Ca2+-ATPase as an example. Nat. Protoc., 2008, 3(11), 1782-1795.[http://dx.doi.org/10.1038/nprot.2008.177] [PMID: 18974737]
[47]
Gorka, J.; Rohmer, M.; Bornemann, S.; Papasotiriou, D.G.; Baeumlisberger, D.; Arrey, T.N.; Bahr, U.; Karas, M. Perfusion reversed-phase high-performance liquid chromatography for protein separation from detergent-containing solutions: an alternative to gel-based approaches. Anal. Biochem., 2012, 424(2), 97-107.[http://dx.doi.org/10.1016/j.ab.2012.02.021] [PMID: 22370273]
[48]
Ehsan, M.; Du, Y.; Mortensen, J.S.; Hariharan, P.; Qu, Q.; Ghani, L.; Das, M.; Grethen, A.; Byrne, B.; Skiniotis, G.; Keller, S.; Loland, C.J.; Guan, L.; Kobilka, B.K.; Chae, P.S. Self-assembly behavior and application of terphenyl-cored trimaltosides for membrane-protein studies: impact of detergent hydrophobic group geometry on protein stability. Chemistry, 2019, 25(49), 11545-11554.[http://dx.doi.org/10.1002/chem.201902468] [PMID: 31243822]
[49]
Anandan, A.; Vrielink, A. Detergents in membrane protein purification and crystallisation. Adv. Exp. Med. Biol., 2016, 922, 13-28.[http://dx.doi.org/10.1007/978-3-319-35072-1_2] [PMID: 27553232]
[50]
Bechara, C.; Bolbach, G.; Bazzaco, P.; Sharma, K.S.; Durand, G.; Popot, J.L.; Zito, F.; Sagan, S. MALDI-TOF mass spectrometry analysis of amphipol-trapped membrane proteins. Anal. Chem., 2012, 84(14), 6128-6135.[http://dx.doi.org/10.1021/ac301035r] [PMID: 22703540]
[51]
Burton-Smith, R.N.; Watanabe, A.; Tokutsu, R.; Song, C.; Murata, K.; Minagawa, J. Structural determination of the large photosystem II-light-harvesting complex II supercomplex of Chlamydomonas reinhardtii using nonionic amphipol. J. Biol. Chem., 2019, 294(41), 15003-15013.[http://dx.doi.org/10.1074/jbc.RA119.009341] [PMID: 31420447]
[52]
Hagn, F.; Nasr, M.L.; Wagner, G. Assembly of phospholipid nanodiscs of controlled size for structural studies of membrane proteins by NMR. Nat. Protoc., 2018, 13(1), 79-98.[http://dx.doi.org/10.1038/nprot.2017.094] [PMID: 29215632]
[53]
Yeh, V.; Lee, T.Y.; Chen, C.W.; Kuo, P.C.; Shiue, J.; Chu, L.K.; Yu, T.Y. Highly efficient transfer of 7TM membrane protein from native membrane to covalently circularized nanodisc. Sci. Rep., 2018, 8(1), 13501.[http://dx.doi.org/10.1038/s41598-018-31925-1] [PMID: 30201976]

Rights & Permissions Print Cite
© 2025 Bentham Science Publishers | Privacy Policy
Request history8.2.27PHP Version317msRequest Duration28MBMemory UsageGET article/{article_id}/{param?}Route
  • warninglog[19:54:55] LOG.warning: Optional parameter $article_id declared before required parameter $request i...
  • warninglog[19:54:56] LOG.warning: Function strftime() is deprecated in /home/alphaeurekaselec/public_html/stor...
  • warninglog[19:54:56] LOG.warning: trim(): Passing null to parameter #1 ($string) of type string is deprecated ...
  • warninglog[19:54:56] LOG.warning: Function strftime() is deprecated in /home/alphaeurekaselec/public_html/stor...
  • Booting (123ms)time
  • Application (194ms)time
  • 1 x Application (61.2%)
    194ms
    1 x Booting (38.8%)
    123ms
    14 templates were rendered
    • articlearticle.blade.php#?blade
    • includes.article_top_bannerarticle_top_banner.blade.php#?blade
    • includes.article_referencearticle_reference.blade.php#?blade
    • export_citation_formexport_citation_form.blade.php#?blade
    • includes.article_bottom_bannerarticle_bottom_banner.blade.php#?blade
    • includes.inc_related_journalsinc_related_journals.blade.php#?blade
    • includes.inc_related_ebooksinc_related_ebooks.blade.php#?blade
    • includes.journal_rightmenujournal_rightmenu.blade.php#?blade
    • includes.journal_right_bannerjournal_right_banner.blade.php#?blade
    • layouts.appapp.blade.php#?blade
    • layouts._login_access_login_access.blade.php#?blade
    • layouts._header_header.blade.php#?blade
    • layouts._main_menu_main_menu.blade.php#?blade
    • layouts._footer_menu_footer_menu.blade.php#?blade
    uri
    GET article/{article_id}/{param?}
    middleware
    web, check_user_access
    controller
    App\Http\Controllers\ArticleController@GetArticleDetails
    namespace
    App\Http\Controllers
    where
    as
    article
    file
    app/Http/Controllers/ArticleController.php:1974-2510
    36 statements were executed, 4 of which were duplicates, 32 unique. Show only duplicated123ms
    • Connection Establishedalphaeurekaselec_live_10_06_2022UserAccessCheck.php#24
      Backtrace
      • 9. middleware::check_user_access:24
      • 10. vendor/laravel/framework/src/Illuminate/Pipeline/Pipeline.php:183
      • 11. vendor/laravel/framework/src/Illuminate/Routing/Middleware/SubstituteBindings.php:50
      • 12. vendor/laravel/framework/src/Illuminate/Pipeline/Pipeline.php:183
      • 13. vendor/laravel/framework/src/Illuminate/Foundation/Http/Middleware/VerifyCsrfToken.php:88
    • INSERT INTO visitor_ip_address (ip_address,visitor_agent,visitor_count, created_on,updated_on) VALUES ('309686912','',1,1740513295,1740513295) ON DUPLICATE KEY UPDATE visitor_count=visitor_count + 1, updated_on=1740513295
      860μsalphaeurekaselec_live_10_06_2022UserAccessCheck.php#24
      Backtrace
      • 11. middleware::check_user_access:24
      • 12. vendor/laravel/framework/src/Illuminate/Pipeline/Pipeline.php:183
      • 13. vendor/laravel/framework/src/Illuminate/Routing/Middleware/SubstituteBindings.php:50
      • 14. vendor/laravel/framework/src/Illuminate/Pipeline/Pipeline.php:183
      • 15. vendor/laravel/framework/src/Illuminate/Foundation/Http/Middleware/VerifyCsrfToken.php:88
    • INSERT INTO visitor_ip_address_detail (ip_address,visitor_url,visitor_count, created_on,updated_on) VALUES ('309686912','/article/115351',1,1740513295,1740513295)
      140μsalphaeurekaselec_live_10_06_2022UserAccessCheck.php#27
      Backtrace
      • 11. middleware::check_user_access:27
      • 12. vendor/laravel/framework/src/Illuminate/Pipeline/Pipeline.php:183
      • 13. vendor/laravel/framework/src/Illuminate/Routing/Middleware/SubstituteBindings.php:50
      • 14. vendor/laravel/framework/src/Illuminate/Pipeline/Pipeline.php:183
      • 15. vendor/laravel/framework/src/Illuminate/Foundation/Http/Middleware/VerifyCsrfToken.php:88
    • (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_journal_volume_trail WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_journal_volume_corporate WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_journal_volume_token WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_ebook_volume_trail WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_ebook_volume_corporate WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_ebook_volume_token WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_issue_trail WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_issue_corporate WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_issue_token WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_article_trail WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_article_corporate WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_article_token WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_chapter_trail WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_chapter_corporate WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_chapter_token WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) union (select `user_id`, `to_ip`, `from_ip` from (select user_id, to_ip, from_ip FROM user_access_disease_corporate WHERE to_ip >= 309686912 ORDER BY to_ip ASC LIMIT 1) as `tjv` where `from_ip` <= 309686912) limit 1
      12.39msalphaeurekaselec_live_10_06_2022UserAccess.php#784
      Bindings
      • 0: 309686912
      • 1: 309686912
      • 2: 309686912
      • 3: 309686912
      • 4: 309686912
      • 5: 309686912
      • 6: 309686912
      • 7: 309686912
      • 8: 309686912
      • 9: 309686912
      • 10: 309686912
      • 11: 309686912
      • 12: 309686912
      • 13: 309686912
      • 14: 309686912
      • 15: 309686912
      Backtrace
      • 14. app/Models/UserAccess/UserAccess.php:784
      • 15. middleware::check_user_access:114
      • 16. vendor/laravel/framework/src/Illuminate/Pipeline/Pipeline.php:183
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Middleware/SubstituteBindings.php:50
      • 18. vendor/laravel/framework/src/Illuminate/Pipeline/Pipeline.php:183
    • SELECT at.type_name, p.copyright, p.publisher_location,m.month,y.year, a.publish_on, a.nid article_nid,a.article_id, a.pmid, a.issue_id, a.title, a.abstract,a.podcast,a.altmetric_score,a.altmetric_url,a.kudos_url, a.page_count, a.first_page, a.elocator, a.last_page, i.publication_date, a.file_name, a.file_path, a.doi, a.is_epub,a.epub_type, a.volume_id, a.is_uploaded, a.editor_choice, a.is_epub, j.image_file_name, a.article_no, a.art_type AS article_type, a.supplementary_filename,a.html_filename,a.prc_filename,a.epub_filename, a.graph_abs_lg_filename, a.graph_abs_sm_filename, a.oa_image_lg_filename, a.oa_image_sm_filename, j.journal_banner,a.animated_abstract,a.erratum_in,a.erratum_for, (CASE WHEN a.issue_id =0 THEN j.epub_price ELSE j.price END) AS sell_price, j.epub_price, j.nid AS jour_nid, j.title AS journal_title, j.issn, j.eissn,j.journal_id, j.subtitle AS jour_subtitle, v.volume_id,v.volume_name AS volume, v.year_id, i.title AS issue, i.month_id ,i.type issue_type, j.subtitle,j.formerly_title, ac.subtitle AS epub_title,a.crossmark_enabled,a.text_mining_urls,a.license_urls,a.created, a.receivedate, a.revisedate, a.acceptdate,js.js_title FROM article a LEFT JOIN article_section ac ON a.is_epub = ac.art_sec_id LEFT JOIN journal_section js ON a.js_id = js.js_id LEFT JOIN article_type at ON a.art_type = at.art_type_id LEFT JOIN issue i ON a.issue_id = i.issue_id LEFT JOIN month m ON m.id = i.month_id LEFT JOIN volume v ON v.volume_id = a.volume_id INNER JOIN year y ON y.id = v.year_id INNER JOIN journal j ON j.journal_id = a.journal_id INNER JOIN publisher p ON p.publisher_id = j.publisher_id WHERE a.article_id=115351
      640μsalphaeurekaselec_live_10_06_2022Article.php#408
      Backtrace
      • 11. app/Models/Article.php:408
      • 12. app/Http/Controllers/ArticleController.php:2515
      • 13. app/Http/Controllers/ArticleController.php:2035
      • 14. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 15. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select * from `journal` where `journal_id` = 51 limit 1
      740μsalphaeurekaselec_live_10_06_2022Journal.php#64
      Bindings
      • 0: 51
      Backtrace
      • 14. app/Models/Journal.php:64
      • 15. app/Http/Controllers/ArticleController.php:2516
      • 16. app/Http/Controllers/ArticleController.php:2035
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 18. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select count(*) as aggregate from (select j.nid as journal_nid,j.flyer_link,j.title ,j.issn,j.eissn,j.keyword, j.journal_id as journal_id, j.subtitle, j.image_file_name,j.description,j.journal_insight_url,j.doi from `subject_journal` as `sj` inner join `journal` as `j` on `j`.`journal_id` = `sj`.`journal_id` where j.journal_status=1 and subject_id IN (SELECT subject_id FROM subject_journal WHERE journal_id=51) and sj.journal_id!=51 group by `j`.`nid`) as `aggregate_table`
      790μsalphaeurekaselec_live_10_06_2022Subject.php#200
      Backtrace
      • 16. app/Models/Subject.php:200
      • 17. app/Http/Controllers/ArticleController.php:2517
      • 18. app/Http/Controllers/ArticleController.php:2035
      • 19. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 20. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select j.nid as journal_nid,j.flyer_link,j.title ,j.issn,j.eissn,j.keyword, j.journal_id as journal_id, j.subtitle, j.image_file_name,j.description,j.journal_insight_url,j.doi from `subject_journal` as `sj` inner join `journal` as `j` on `j`.`journal_id` = `sj`.`journal_id` where j.journal_status=1 and subject_id IN (SELECT subject_id FROM subject_journal WHERE journal_id=51) and sj.journal_id!=51 group by `j`.`nid` order by `j`.`journal_id` asc limit 10 offset 0
      650μsalphaeurekaselec_live_10_06_2022Subject.php#200
      Backtrace
      • 14. app/Models/Subject.php:200
      • 15. app/Http/Controllers/ArticleController.php:2517
      • 16. app/Http/Controllers/ArticleController.php:2035
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 18. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select count(*) as aggregate from (select v.nid as volume_id,e.nid,e.title,v.issn,v.eissn,v.isbn,v.eisbn,e.ebook_id, v.image_file_name,v.file_path,v.flyer_link,v.ebook_volume_id, v.volume_name,v.doi,v.year_id,v.introduction from `subject_ebook` as `se` inner join `ebook` as `e` on `e`.`ebook_id` = `se`.`ebook_id` inner join `ebook_volume` as `v` on `v`.`ebook_id` = `se`.`ebook_id` inner join `year` as `y` on `y`.`id` = `v`.`year_id` where subject_id IN (SELECT subject_id FROM subject_journal WHERE journal_id=51) and v.ebook_status='1' and y.year > 2020 group by `v`.`nid`) as `aggregate_table`
      2.58msalphaeurekaselec_live_10_06_2022Subject.php#220
      Backtrace
      • 16. app/Models/Subject.php:220
      • 17. app/Http/Controllers/ArticleController.php:2518
      • 18. app/Http/Controllers/ArticleController.php:2035
      • 19. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 20. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select v.nid as volume_id,e.nid,e.title,v.issn,v.eissn,v.isbn,v.eisbn,e.ebook_id, v.image_file_name,v.file_path,v.flyer_link,v.ebook_volume_id, v.volume_name,v.doi,v.year_id,v.introduction from `subject_ebook` as `se` inner join `ebook` as `e` on `e`.`ebook_id` = `se`.`ebook_id` inner join `ebook_volume` as `v` on `v`.`ebook_id` = `se`.`ebook_id` inner join `year` as `y` on `y`.`id` = `v`.`year_id` where subject_id IN (SELECT subject_id FROM subject_journal WHERE journal_id=51) and v.ebook_status='1' and y.year > 2020 group by `v`.`nid` order by `v`.`ebook_volume_id` desc limit 10 offset 0
      2.08msalphaeurekaselec_live_10_06_2022Subject.php#220
      Backtrace
      • 14. app/Models/Subject.php:220
      • 15. app/Http/Controllers/ArticleController.php:2518
      • 16. app/Http/Controllers/ArticleController.php:2035
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 18. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select * from `keywords` where `article_id` = '115351'
      1.04msalphaeurekaselec_live_10_06_2022Keywords.php#43
      Bindings
      • 0: 115351
      Backtrace
      • 13. app/Models/Keywords.php:43
      • 14. app/Http/Controllers/ArticleController.php:2519
      • 15. app/Http/Controllers/ArticleController.php:2035
      • 16. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 17. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select * from `article_citation` where `article_id` = '115351'
      2.23msalphaeurekaselec_live_10_06_2022Article.php#1570
      Bindings
      • 0: 115351
      Backtrace
      • 13. app/Models/Article.php:1570
      • 14. app/Http/Controllers/ArticleController.php:2520
      • 15. app/Http/Controllers/ArticleController.php:2035
      • 16. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 17. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select CONCAT_WS(' ',au.first_name, ' ', ifnull(au.initials,''), ' ', au.last_name) as authors, CONCAT(au.last_name, IFNULL(CONCAT(' ', au.initials), ''), ' ', au.first_name) AS authorsRIS, CONCAT_WS(' ',au.last_name, ' ', ifnull(au.initials,''), ' ', au.first_name) as authorsCiteAs, CONCAT_WS(' ',au.first_name, IFNULL(CONCAT(' ', au.initials), ''), IFNULL(CONCAT(' ', au.last_name), '')) as authorsmodal , `au`.*, `af`.`ror_id`, `af`.`institution`, `af`.`department`, `af`.`country`, `af`.`city`, `af`.`address`, (GROUP_CONCAT(TRIM(BOTH ', ' FROM CONCAT(ifnull(concat(af.institution,','),''), ifnull(concat(af.department,','),''), ifnull(concat(af.address,','),''), ifnull(concat(af.city,','),''), ifnull(concat(af.country,','),''), ifnull(concat(af.postal_code,','),''), ifnull(concat(af.phone,','),''), ifnull(concat(af.fax,','),''))) SEPARATOR '|')) as `author_affiliation`, (GROUP_CONCAT(TRIM(BOTH ', ' FROM CONCAT(ifnull(concat(af.web_view,','),''))) SEPARATOR '|')) as `web_view` from `author` as `au` left join `author_affiliation` as `af` on `au`.`author_id` = `af`.`author_id` where `au`.`article_id` = '115351' group by `au`.`author_id` order by `au`.`article_id` asc, `au`.`sequence` asc
      5.34msalphaeurekaselec_live_10_06_2022Author.php#86
      Bindings
      • 0: 115351
      Backtrace
      • 13. app/Models/Author.php:86
      • 14. app/Http/Controllers/ArticleController.php:2533
      • 15. app/Http/Controllers/ArticleController.php:2035
      • 16. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 17. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select `bo`.* from `bundle_offer` as `bo` where `bo`.`bundle_status` = 'A' order by `bo`.`bundle_id` desc
      230μsalphaeurekaselec_live_10_06_2022BundleOffer.php#57
      Bindings
      • 0: A
      Backtrace
      • 13. app/Models/BundleOffer.php:57
      • 14. app/Http/Controllers/ArticleController.php:2535
      • 15. app/Http/Controllers/ArticleController.php:2035
      • 16. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 17. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
    • select `subtitle` from `journal` where `continues_publication_journal` = 1
      410μsalphaeurekaselec_live_10_06_2022Article.php#1960
      Bindings
      • 0: 1
      Backtrace
      • 13. app/Models/Article.php:1960
      • 14. app/Http/Controllers/ArticleController.php:2062
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • Select j.subtitle, v.volume_id as volume_id,v.year_id,y.year as year,v.volume_name, v.total_no_of_issues, i.issue_id as issue_id,i.title as issue, i.publication_date as issue_pub_date from journal j join volume v on j.journal_id = v.journal_id join issue i on v.volume_id = i.volume_id join article a on a.issue_id = i.issue_id join year y on v.year_id = y.id where j.journal_id = 51 and i.is_uploaded = 1 order by v.volume_name+0 desc, i.title+0 desc limit 1
      27.54msalphaeurekaselec_live_10_06_2022Issue.php#298
      Backtrace
      • 11. app/Models/Issue.php:298
      • 12. app/Http/Controllers/ArticleController.php:2068
      • 13. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 14. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select a.nid,a.publish_on, a.pmid,a.article_id,a.doi,j.journal_id,j.subtitle,v.volume_name,a.volume_id as volume_id, a.issue_id,a.title,first_page, last_page, page_count,abstract,ifnull(is_abstract,"y"),asec.title as article_section,at.type_name as article_type_name, a.article_id as ArticleID ,is_epub,epub_price,epub_type,a.created, y.year from `article` as `a` left join `article_section` as `asec` on `asec`.`art_sec_id` = `a`.`is_epub` left join `article_type` as `at` on `at`.`art_type_id` = `a`.`art_type` left join `journal` as `j` on `j`.`journal_id` = `a`.`journal_id` inner join `volume` as `v` on `a`.`volume_id` = `v`.`volume_id` inner join `year` as `y` on `v`.`year_id` = `y`.`id` where a.article_status != "W" and a.is_epub = 1 and a.journal_id = 51 and a.is_uploaded = 1 group by `a`.`article_id` order by `a`.`publish_on` desc
      20.02msalphaeurekaselec_live_10_06_2022Article.php#528
      Backtrace
      • 13. app/Models/Article.php:528
      • 14. app/Http/Controllers/ArticleController.php:2070
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select a.nid,a.publish_on, a.pmid,a.article_id,a.doi,j.journal_id,j.subtitle,v.volume_name,a.volume_id as volume_id, a.issue_id,a.title,first_page, last_page, page_count,abstract,ifnull(is_abstract,"y"),asec.title as article_section,at.type_name as article_type_name, a.article_id as ArticleID ,is_epub,epub_price,epub_type,a.created, y.year from `article` as `a` left join `article_section` as `asec` on `asec`.`art_sec_id` = `a`.`is_epub` left join `article_type` as `at` on `at`.`art_type_id` = `a`.`art_type` left join `journal` as `j` on `j`.`journal_id` = `a`.`journal_id` inner join `volume` as `v` on `a`.`volume_id` = `v`.`volume_id` inner join `year` as `y` on `v`.`year_id` = `y`.`id` where a.article_status != "W" and a.is_epub = 4 and a.journal_id = 51 and a.is_uploaded = 1 group by `a`.`article_id` order by `a`.`publish_on` desc
      14.78msalphaeurekaselec_live_10_06_2022Article.php#528
      Backtrace
      • 13. app/Models/Article.php:528
      • 14. app/Http/Controllers/ArticleController.php:2071
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select pdf, html, epub, prc from `article_metrics` where `article_id` = 115351 limit 1
      210μsalphaeurekaselec_live_10_06_2022Article.php#744
      Bindings
      • 0: 115351
      Backtrace
      • 14. app/Models/Article.php:744
      • 15. app/Http/Controllers/ArticleController.php:2091
      • 16. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 17. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 18. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select * from content_by_diseases where content_id = 115351 and content_type = 'article' order by status desc limit 1
      250μsalphaeurekaselec_live_10_06_2022Article.php#751
      Backtrace
      • 11. app/Models/Article.php:751
      • 12. app/Http/Controllers/ArticleController.php:2094
      • 13. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 14. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select * from `node_meta` where `nid` = 192863 limit 1
      190μsalphaeurekaselec_live_10_06_2022Meta.php#30
      Bindings
      • 0: 192863
      Backtrace
      • 14. app/Models/Meta.php:30
      • 15. app/Http/Controllers/ArticleController.php:2101
      • 16. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 17. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 18. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select `bo`.* from `bundle_offer` as `bo` where `bo`.`bundle_status` = 'A' order by `bo`.`bundle_id` desc
      190μsalphaeurekaselec_live_10_06_2022BundleOffer.php#57
      Bindings
      • 0: A
      Backtrace
      • 13. app/Models/BundleOffer.php:57
      • 14. app/Http/Controllers/ArticleController.php:2108
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select * from `tbl_ht_submission` as `h` where `h`.`subtitle` = 'PPL' and (h.proposal_closing_date = 0 OR TO_DAYS(FROM_UNIXTIME(h.proposal_closing_date)) >= TO_DAYS(NOW())) order by `h`.`manuscript` asc
      230μsalphaeurekaselec_live_10_06_2022Article.php#2025
      Bindings
      • 0: PPL
      Backtrace
      • 13. app/Models/Article.php:2025
      • 14. app/Http/Controllers/ArticleController.php:2112
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select count(*) as total_count, content_type_access_id as access_type from journal_access where content_type_id = '115351' and content_type = 'a' and ( (from_date <= CURDATE() and to_date >= CURDATE() and perpetual= 0) or (from_date is null and to_date is null and perpetual= 1) )
      330μsalphaeurekaselec_live_10_06_2022ContentAccess.php#647
      Backtrace
      • 11. app/Models/ContentAccess.php:647
      • 12. app/Http/Controllers/ArticleController.php:2186
      • 13. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 14. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select count(*) as total_count, content_type_access_id as access_type from journal_access where content_type_id = '10840' and content_type = 'i' and ( (from_date <= CURDATE() and to_date >= CURDATE() and perpetual= 0) or (from_date is null and to_date is null and perpetual= 1) )
      260μsalphaeurekaselec_live_10_06_2022ContentAccess.php#647
      Backtrace
      • 11. app/Models/ContentAccess.php:647
      • 12. app/Http/Controllers/ArticleController.php:2207
      • 13. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 14. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select count(*) as total_count, content_type_access_id as access_type from journal_access where content_type_id = '2944' and content_type = 'v' and ( (from_date <= CURDATE() and to_date >= CURDATE() and perpetual= 0) or (from_date is null and to_date is null and perpetual= 1) )
      250μsalphaeurekaselec_live_10_06_2022ContentAccess.php#647
      Backtrace
      • 11. app/Models/ContentAccess.php:647
      • 12. app/Http/Controllers/ArticleController.php:2231
      • 13. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 14. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Trial Access" as access_base_text, CASE j.type WHEN 111 THEN true ELSE false END as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"volume" as access_level,"" as issue_id,"" as article_id from `user_access_journal_volume_trail` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 309686912 and j.to_ip >=309686912 )) and ((j.from_date <= 1740513296 and j.to_date >= 1740513296 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.volume_id = 2944) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Subscribed" as access_base_text, false as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"volume" as access_level,"" as issue_id,"" as article_id from `user_access_journal_volume_corporate` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 309686912 and j.to_ip >=309686912 )) and ((j.from_date <= 1740513296 and j.to_date >= 1740513296 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.volume_id = 2944) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Token Based Access" as access_base_text, false as terms_popup, "R" as content_download_type, j.uc_id as restricted_user_access_key, "v" as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"volume" as access_level,"" as issue_id,"" as article_id from `user_access_journal_volume_token` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 309686912 and j.to_ip >=309686912 )) and ((j.from_date <= 1740513296 and j.to_date >= 1740513296 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.volume_id = 2944)
      21.59msalphaeurekaselec_live_10_06_2022UserAccess.php#1266
      Backtrace
      • 13. app/Models/UserAccess/UserAccess.php:1266
      • 14. app/Http/Controllers/ArticleController.php:2272
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Trial Access" as access_base_text, CASE j.type WHEN 111 THEN true ELSE false END as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"issue" as access_level,j.issue_id,"" as article_id from `user_access_issue_trail` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 309686912 and j.to_ip >=309686912 )) and ((j.from_date <= 1740513296 and j.to_date >= 1740513296 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.issue_id = 10840) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Subscribed" as access_base_text, false as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"issue" as access_level,j.issue_id,"" as article_id from `user_access_issue_corporate` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 309686912 and j.to_ip >=309686912 )) and ((j.from_date <= 1740513296 and j.to_date >= 1740513296 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.issue_id = 10840) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Token Based Access" as access_base_text, false as terms_popup, "R" as content_download_type, j.uc_id as restricted_user_access_key, "v" as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"issue" as access_level,j.issue_id,"" as article_id from `user_access_issue_token` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 309686912 and j.to_ip >=309686912 )) and ((j.from_date <= 1740513296 and j.to_date >= 1740513296 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.issue_id = 10840)
      600μsalphaeurekaselec_live_10_06_2022UserAccess.php#1266
      Backtrace
      • 13. app/Models/UserAccess/UserAccess.php:1266
      • 14. app/Http/Controllers/ArticleController.php:2297
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Trial Access" as access_base_text, CASE j.type WHEN 111 THEN true ELSE false END as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"article" as access_level, j.issue_id, j.article_id from `user_access_article_trail` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 309686912 and j.to_ip >=309686912 )) and ((j.from_date <= 1740513296 and j.to_date >= 1740513296 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.article_id = 115351) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Subscribed" as access_base_text, false as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"article" as access_level, j.issue_id, j.article_id from `user_access_article_corporate` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 309686912 and j.to_ip >=309686912 )) and ((j.from_date <= 1740513296 and j.to_date >= 1740513296 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.article_id = 115351) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Token Based Access" as access_base_text, false as terms_popup, "R" as content_download_type, j.uc_id as restricted_user_access_key, "v" as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"article" as access_level, j.issue_id, j.article_id from `user_access_article_token` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 309686912 and j.to_ip >=309686912 )) and ((j.from_date <= 1740513296 and j.to_date >= 1740513296 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.article_id = 115351)
      550μsalphaeurekaselec_live_10_06_2022UserAccess.php#1266
      Backtrace
      • 13. app/Models/UserAccess/UserAccess.php:1266
      • 14. app/Http/Controllers/ArticleController.php:2329
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select * from `article` where `first_page` < '972' and is_uploaded = 1 and issue_id = 10840 order by cast(first_page as SIGNED) desc limit 1
      460μsalphaeurekaselec_live_10_06_2022Article.php#1819
      Bindings
      • 0: 972
      Backtrace
      • 14. app/Models/Article.php:1819
      • 15. app/Http/Controllers/ArticleController.php:2470
      • 16. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 17. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 18. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select * from `article` where `last_page` > '982' and is_uploaded = 1 and issue_id = 10840 order by cast(first_page as SIGNED) asc limit 1
      380μsalphaeurekaselec_live_10_06_2022Article.php#1831
      Bindings
      • 0: 982
      Backtrace
      • 14. app/Models/Article.php:1831
      • 15. app/Http/Controllers/ArticleController.php:2472
      • 16. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 17. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 18. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select `language_id` from `multilanguage` where `short_code` = 'en'
      150μsalphaeurekaselec_live_10_06_2022ArticleController.php#2497
      Bindings
      • 0: en
      Backtrace
      • 14. app/Http/Controllers/ArticleController.php:2497
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
      • 18. vendor/laravel/framework/src/Illuminate/Routing/Route.php:206
    • select `source_lang_label`, `target_lang_label` from `multilanguage_labels` where `target_lang_id` = 1
      170μsalphaeurekaselec_live_10_06_2022MultiLanguage.php#23
      Bindings
      • 0: 1
      Backtrace
      • 13. app/Models/MultiLanguage.php:23
      • 14. app/Http/Controllers/ArticleController.php:2498
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select `id` from `multilanguage_article` where `article_id` = 115351 limit 1
      2.33msalphaeurekaselec_live_10_06_2022ArticleController.php#2506
      Bindings
      • 0: 115351
      Backtrace
      • 16. app/Http/Controllers/ArticleController.php:2506
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 18. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 19. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
      • 20. vendor/laravel/framework/src/Illuminate/Routing/Route.php:206
    • select b.banner_link, b.banner_name, b.banner_script, b.banner_remarks, b.banner_file_name, b.banner_placement, bd.content_type, bd.content_id from `tbl_banner` as `b` inner join `tbl_banner_detail` as `bd` on `bd`.`banner_id` = `b`.`banner_id` inner join `users` as `u` on `u`.`id` = `b`.`updated_by` inner join `journal` as `j` on `j`.`journal_id` = `bd`.`content_id` where `b`.`banner_journal` = 1 and `bd`.`content_type` = 'J' and `bd`.`content_id` = 51 and `b`.`banner_from_date` <= 1740513296 and `b`.`banner_to_date` >= 1740513296 and `b`.`banner_status` = 'A' order by `b`.`banner_id` desc
      1.11msalphaeurekaselec_live_10_06_2022Banner.php#95
      Bindings
      • 0: 1
      • 1: J
      • 2: 51
      • 3: 1740513296
      • 4: 1740513296
      • 5: A
      Backtrace
      • 13. app/Models/Banner.php:95
      • 14. app/Http/helpers.php:404
      • 17. vendor/laravel/framework/src/Illuminate/Filesystem/Filesystem.php:124
      • 18. vendor/laravel/framework/src/Illuminate/View/Engines/PhpEngine.php:58
      • 19. vendor/laravel/framework/src/Illuminate/View/Engines/CompilerEngine.php:73
    • select b.banner_link, b.banner_name, b.banner_script, b.banner_remarks, b.banner_file_name, b.banner_placement, bd.content_type, bd.content_id from `tbl_banner` as `b` inner join `tbl_banner_detail` as `bd` on `bd`.`banner_id` = `b`.`banner_id` inner join `users` as `u` on `u`.`id` = `b`.`updated_by` inner join `journal` as `j` on `j`.`journal_id` = `bd`.`content_id` where `b`.`banner_journal` = 1 and `bd`.`content_type` = 'J' and `bd`.`content_id` = 51 and `b`.`banner_from_date` <= 1740513296 and `b`.`banner_to_date` >= 1740513296 and `b`.`banner_status` = 'A' order by `b`.`banner_id` desc
      1.14msalphaeurekaselec_live_10_06_2022Banner.php#95
      Bindings
      • 0: 1
      • 1: J
      • 2: 51
      • 3: 1740513296
      • 4: 1740513296
      • 5: A
      Backtrace
      • 13. app/Models/Banner.php:95
      • 14. app/Http/helpers.php:404
      • 17. vendor/laravel/framework/src/Illuminate/Filesystem/Filesystem.php:124
      • 18. vendor/laravel/framework/src/Illuminate/View/Engines/PhpEngine.php:58
      • 19. vendor/laravel/framework/src/Illuminate/View/Engines/CompilerEngine.php:73
    • select count(*) as aggregate from `uc_cart_products` where cart_id='aabdd3d3bd1d69bea540403e29fa8a79'
      200μsalphaeurekaselec_live_10_06_2022Cart.php#271
      Backtrace
      • 15. app/Models/Cart.php:271
      • 16. view::layouts._header:260
      • 18. vendor/laravel/framework/src/Illuminate/Filesystem/Filesystem.php:124
      • 19. vendor/laravel/framework/src/Illuminate/View/Engines/PhpEngine.php:58
      • 20. vendor/laravel/framework/src/Illuminate/View/Engines/CompilerEngine.php:73
    App\Models\MultiLanguage
    1MultiLanguage.php#?
        _token
        eWj09NyimllUJnznr6r4Q7BHLUbD0XgkqLDHNN7F
        uc_cart_id
        aabdd3d3bd1d69bea540403e29fa8a79
        _previous
        array:1 [ "url" => "http://alpha.eurekaselect.com/article/115351" ]
        _flash
        array:2 [ "old" => [] "new" => [] ]
        PHPDEBUGBAR_STACK_DATA
        []
        path_info
        /article/115351
        status_code
        200
        
        status_text
        OK
        format
        html
        content_type
        text/html; charset=UTF-8
        request_query
        []
        
        request_request
        []
        
        request_headers
        0 of 0
        array:8 [ "accept-encoding" => array:1 [ 0 => "gzip, deflate" ] "accept" => array:1 [ 0 => "text/html,application/xhtml+xml,application/xml;q=0.9,image/avif,image/webp,image/apng,*/*;q=0.8,application/signed-exchange;v=b3;q=0.7" ] "user-agent" => array:1 [ 0 => "Mozilla/5.0 AppleWebKit/537.36 (KHTML, like Gecko; compatible; ClaudeBot/1.0; +claudebot@anthropic.com)" ] "upgrade-insecure-requests" => array:1 [ 0 => "1" ] "cache-control" => array:1 [ 0 => "no-cache" ] "pragma" => array:1 [ 0 => "no-cache" ] "connection" => array:1 [ 0 => "keep-alive" ] "host" => array:1 [ 0 => "alpha.eurekaselect.com" ] ]
        request_cookies
        []
        
        response_headers
        0 of 0
        array:5 [ "content-type" => array:1 [ 0 => "text/html; charset=UTF-8" ] "cache-control" => array:1 [ 0 => "no-cache, private" ] "date" => array:1 [ 0 => "Tue, 25 Feb 2025 19:54:56 GMT" ] "set-cookie" => array:2 [ 0 => "XSRF-TOKEN=eyJpdiI6IjBsdk12L1poU0dPOTY1MVFVVE5tSnc9PSIsInZhbHVlIjoiN2I0UzRVUC9PRWN1TGhDcHdxeXhBZVd2TE42bkp0eGg2OXgrN2tGVmNWWTFMRUJzVUdtalNVT21hK1NRVC82ZnVuMmpxM1JKU2xtYjRGQ2ZUUWpubHJrQ01rVERDSHlLTnhHbkliMEw1VmcvaWJadUFFSzBWaE9HTjByVHlLN3YiLCJtYWMiOiI2ZmJmOWMxNWZmNzJiM2M4YjUzMjgxMjZjNTY4YWNmNWY0MjliZDZkYzlmYmI0YTJkZDEzZGM0MzI2OTdiZTdjIiwidGFnIjoiIn0%3D; expires=Fri, 28 Feb 2025 07:54:56 GMT; Max-Age=216000; path=/; secure; samesite=laxXSRF-TOKEN=eyJpdiI6IjBsdk12L1poU0dPOTY1MVFVVE5tSnc9PSIsInZhbHVlIjoiN2I0UzRVUC9PRWN1TGhDcHdxeXhBZVd2TE42bkp0eGg2OXgrN2tGVmNWWTFMRUJzVUdtalNVT21hK1NRVC82ZnVuMmpxM" 1 => "alphaeurekaselectnet_session=eyJpdiI6IitjMTlpYXlnN3RSSHNuS2xtNWlOYlE9PSIsInZhbHVlIjoiVGIxY2dMSGFYOHlBRE5JbVVPL2F1Y1J6a1E1d0VpODI0V2ZjR2N2TFhDdkowRmQxTEo1b0E0SGhFTjJPcnVDTUgwNXUxeG5SNFY3eVUwZHJiUm8zNy9KT094K0VuMzdiQnFGNVZLTmF0RUNlV3paYkRZZjhGVUk0TkE2SUIydjIiLCJtYWMiOiJlNGI3ZTNhMGYxYTE1N2M1OThhM2FhZDAwY2EzZTM2MzU5NTIwZGZmNjY2ZDIzMjc5MDkxZDNkYzEwNzUxNzM3IiwidGFnIjoiIn0%3D; path=/; secure; httponly; samesite=laxalphaeurekaselectnet_session=eyJpdiI6IitjMTlpYXlnN3RSSHNuS2xtNWlOYlE9PSIsInZhbHVlIjoiVGIxY2dMSGFYOHlBRE5JbVVPL2F1Y1J6a1E1d0VpODI0V2ZjR2N2TFhDdkowRmQxTEo1b0E0SGh" ] "Set-Cookie" => array:2 [ 0 => "XSRF-TOKEN=eyJpdiI6IjBsdk12L1poU0dPOTY1MVFVVE5tSnc9PSIsInZhbHVlIjoiN2I0UzRVUC9PRWN1TGhDcHdxeXhBZVd2TE42bkp0eGg2OXgrN2tGVmNWWTFMRUJzVUdtalNVT21hK1NRVC82ZnVuMmpxM1JKU2xtYjRGQ2ZUUWpubHJrQ01rVERDSHlLTnhHbkliMEw1VmcvaWJadUFFSzBWaE9HTjByVHlLN3YiLCJtYWMiOiI2ZmJmOWMxNWZmNzJiM2M4YjUzMjgxMjZjNTY4YWNmNWY0MjliZDZkYzlmYmI0YTJkZDEzZGM0MzI2OTdiZTdjIiwidGFnIjoiIn0%3D; expires=Fri, 28-Feb-2025 07:54:56 GMT; path=/; secureXSRF-TOKEN=eyJpdiI6IjBsdk12L1poU0dPOTY1MVFVVE5tSnc9PSIsInZhbHVlIjoiN2I0UzRVUC9PRWN1TGhDcHdxeXhBZVd2TE42bkp0eGg2OXgrN2tGVmNWWTFMRUJzVUdtalNVT21hK1NRVC82ZnVuMmpxM" 1 => "alphaeurekaselectnet_session=eyJpdiI6IitjMTlpYXlnN3RSSHNuS2xtNWlOYlE9PSIsInZhbHVlIjoiVGIxY2dMSGFYOHlBRE5JbVVPL2F1Y1J6a1E1d0VpODI0V2ZjR2N2TFhDdkowRmQxTEo1b0E0SGhFTjJPcnVDTUgwNXUxeG5SNFY3eVUwZHJiUm8zNy9KT094K0VuMzdiQnFGNVZLTmF0RUNlV3paYkRZZjhGVUk0TkE2SUIydjIiLCJtYWMiOiJlNGI3ZTNhMGYxYTE1N2M1OThhM2FhZDAwY2EzZTM2MzU5NTIwZGZmNjY2ZDIzMjc5MDkxZDNkYzEwNzUxNzM3IiwidGFnIjoiIn0%3D; path=/; secure; httponlyalphaeurekaselectnet_session=eyJpdiI6IitjMTlpYXlnN3RSSHNuS2xtNWlOYlE9PSIsInZhbHVlIjoiVGIxY2dMSGFYOHlBRE5JbVVPL2F1Y1J6a1E1d0VpODI0V2ZjR2N2TFhDdkowRmQxTEo1b0E0SGh" ] ]
        session_attributes
        0 of 0
        array:5 [ "_token" => "eWj09NyimllUJnznr6r4Q7BHLUbD0XgkqLDHNN7F" "uc_cart_id" => "aabdd3d3bd1d69bea540403e29fa8a79" "_previous" => array:1 [ "url" => "http://alpha.eurekaselect.com/article/115351" ] "_flash" => array:2 [ "old" => [] "new" => [] ] "PHPDEBUGBAR_STACK_DATA" => [] ]
        ClearShow all
        Date ↕MethodURLData
        #12025-02-25 19:54:56GET/article/115351414361