{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,17]],"date-time":"2026-10-17T07:35:20Z","timestamp":1792222520036,"version":"4.4.0"},"reference-count":63,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,3,24]],"date-time":"2023-03-24T00:00:00Z","timestamp":1679616000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100013286","name":"Specialized Research Fund for the Doctoral Program of Higher Education of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100013286","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p><jats:italic>Neisseria gonorrhoeae<\/jats:italic> is an emerging multidrug resistance pathogen that causes sexually transmitted infections in men and women. The <jats:italic>N. gonorrhoeae<\/jats:italic> has demonstrated an emerging antimicrobial resistance against reported antibiotics, hence fetching the attention of researchers to address this problem. The present <jats:italic>in-silico<\/jats:italic> study aimed to find putative novel drug and vaccine targets against <jats:italic>N. gonorrhoeae<\/jats:italic> infection by the application of bioinformatics approaches. Core genes set of 69\u00a0N<jats:italic>. gonorrhoeae<\/jats:italic> strains was acquired from complete genome sequences. The essential and non-homologous metabolic pathway proteins of <jats:italic>N. gonorrhoeae<\/jats:italic> were identified. Moreover, different bioinformatics databases were used for the downstream analysis. The DrugBank database scanning identified 12 novel drug targets in the prioritized list. They were preferred as drug targets against this bacterium. A viable vaccine is unavailable so far against <jats:italic>N. gonorrhoeae<\/jats:italic> infection. In the current study, two outer-membrane proteins were prioritized as vaccine candidates <jats:italic>via<\/jats:italic> reverse vaccinology approach. The top lead B and T-cells overlapped epitopes were utilized to generate a chimeric vaccine construct combined with immune-modulating adjuvants, linkers, and PADRE sequences. The top ranked prioritized vaccine construct (V7) showed stable molecular interaction with human immune cell receptors as inferred during the molecular docking and MD simulation analyses. Considerable response for immune cells was interpreted by in-silico immune studies. Additional tentative validation is required to ensure the effectiveness of the prioritized vaccine construct against <jats:italic>N. gonorrhoeae<\/jats:italic> infection. The identified proteins can be used for further rational drug and vaccine designing to develop potential therapeutic entities against the multi-drug resistant <jats:italic>N. gonorrhoeae<\/jats:italic>.<\/jats:p>","DOI":"10.3389\/fcimb.2023.1017315","type":"journal-article","created":{"date-parts":[[2023,3,24]],"date-time":"2023-03-24T05:59:40Z","timestamp":1679637580000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":24,"title":["Computer-aided genomic data analysis of drug-resistant Neisseria gonorrhoeae for the Identification of alternative therapeutic targets"],"prefix":"10.3389","volume":"13","author":[{"given":"Aqsa","family":"Qasim","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samavia","family":"Jaan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tehreem Ul","family":"Wara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad","family":"Shehroz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Umar","family":"Nishan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sulaiman","family":"Shams","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohibullah","family":"Shah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suvash Chandra","family":"Ojha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,3,24]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1016\/j.compbiomed.2021.104701","article-title":"Chlamydia trachomatis core genome data mining for promising novel drug targets and chimeric vaccine candidates identification","volume":"136","author":"Aslam","year":"2021","journal-title":"Comput. Biol. Med."},{"key":"B2","doi-asserted-by":"publisher","first-page":"1734","DOI":"10.1016\/j.ygeno.2019.10.009","article-title":"Potential druggable proteins and chimeric vaccine construct prioritization against brucella melitensis from species core genome data","volume":"112","author":"Aslam","year":"2020","journal-title":"Genomics"},{"key":"B3","doi-asserted-by":"publisher","first-page":"678","DOI":"10.1111\/j.1532-5415.1990.tb01429.x","article-title":"Disseminated gonococcal infection","volume":"38","author":"Bar-On","year":"1990","journal-title":"J.\u00a0Am. Geriatr. Soc"},{"key":"B4","doi-asserted-by":"publisher","first-page":"W22","DOI":"10.1093\/nar\/gkw255","article-title":"EDGAR 2.0: an enhanced software platform for comparative gene content analyses","volume":"44","author":"Blom","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"B5","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1471-2105-7-153","article-title":"Predicting population coverage of T-cell epitope-based diagnostics and vaccines","volume":"7","author":"Bui","year":"2006","journal-title":"BMC Bioinf."},{"key":"B6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1471-2105-8-361","article-title":"Development of an epitope conservancy analysis tool to facilitate the design of epitope-based diagnostics and vaccines","volume":"8","author":"Bui","year":"2007","journal-title":"BMC Bioinf."},{"key":"B7","doi-asserted-by":"publisher","first-page":"321","DOI":"10.3390\/ijms16010321","article-title":"tRNAs as antibiotic targets","volume":"16","author":"Chopra","year":"2015","journal-title":"Int. J. Mol. Sci."},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1186\/1752-0509-6-134","article-title":"Computational codon optimization of synthetic gene for protein expression","volume":"6","author":"Chung","year":"2012","journal-title":"BMC Syst. Biol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"1511","DOI":"10.1002\/pro.5560020916","article-title":"Verification of protein structures: Patterns of nonbonded atomic interactions","volume":"2","author":"Colovos","year":"1993","journal-title":"Protein Sci."},{"key":"B10","doi-asserted-by":"publisher","first-page":"846","DOI":"10.1093\/bioinformatics\/btt619","article-title":"AllergenFP: Allergenicity prediction by descriptor fingerprints","volume":"30","author":"Dimitrov","year":"2014","journal-title":"Bioinformatics"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-8-4","article-title":"VaxiJen: A server for prediction of protective antigens, tumour antigens and subunit vaccines","volume":"8","author":"Doytchinova","year":"2007","journal-title":"BMC Bioinf."},{"key":"B12","doi-asserted-by":"publisher","first-page":"243","DOI":"10.1002\/jmr.893","article-title":"Predicting linear b-cell epitopes using string kernels","volume":"21","author":"El-Manzalawy","year":"2008","journal-title":"J. Mol. Recognit."},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2017.00278","article-title":"The immune epitope database and analysis resource in epitope discovery and synthetic vaccine design","volume":"8","author":"Fleri","year":"2017","journal-title":"Front. Immunol."},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/FIMMU.2017.00278","article-title":"The immune epitope database and analysis resource in epitope discovery and synthetic vaccine design","author":"Fleri","year":"2017","journal-title":"Front. Immunol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"571","DOI":"10.1385\/1592598900","article-title":"The proteomics protocols handbook","author":"Gasteiger","year":"2005","journal-title":"Proteomics Protoc. Handb."},{"key":"B16","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1371\/journal.pone.0142563","article-title":"Improving multi-epitope long peptide vaccine potency by using a strategy that enhances CD4+ T help in BALB\/c mice","volume":"10","author":"Ghaffari-Nazari","year":"2015","journal-title":"PloS One"},{"key":"B17","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1475-2859-4-14","article-title":"Improvement of escherichia coli production strains by modification of the phosphoenolpyruvate:sugar phosphotransferase system","volume":"4","author":"Gosset","year":"2005","journal-title":"Microb. Cell Fact."},{"key":"B18","doi-asserted-by":"publisher","first-page":"212","DOI":"10.1128\/AAC.01310-13","article-title":"ARG-annot, a new bioinformatic tool to discover antibiotic resistance genes in bacterial genomes","volume":"58","author":"Gupta","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"B19","doi-asserted-by":"publisher","first-page":"219","DOI":"10.1016\/j.micpath.2018.09.033","article-title":"Reverse vaccinology and subtractive genomics-based putative vaccine targets identification for burkholderia pseudomallei Bp1651","volume":"125","author":"Hizbullah","year":"2018","journal-title":"Microb. Pathog."},{"key":"B20","doi-asserted-by":"publisher","first-page":"3783714","DOI":"10.1155\/2017\/3783714","article-title":"Application of the subtractive genomics and molecular docking analysis for the identification of novel putative drug targets against salmonella enterica subsp. enterica serovar poona","volume":"2017","author":"Hossain","year":"2017","journal-title":"Biomed Res. Int."},{"key":"B21","doi-asserted-by":"publisher","first-page":"W436","DOI":"10.1093\/nar\/gkv462","article-title":"PockDrug-server: A new web server for predicting pocket druggability on holo and apo proteins","volume":"43","author":"Hussein","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.3389\/FMICB.2022.971263","article-title":"Multi-epitope chimeric vaccine designing and novel drug targets prioritization against multi-drug resistant staphylococcus pseudintermedius","volume":"13","author":"Jaan","year":"2022","journal-title":"Front. Microbiol."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3390\/medicines8020011","article-title":"Sexually transmitted neisseria gonorrhoeae infections\u2013update on drug treatment and vaccine development","volume":"8","author":"Jefferson","year":"2021","journal-title":"Medicines"},{"key":"B24","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1093\/nar\/gkn201","article-title":"NCBI BLAST: a better web interface","volume":"36","author":"Johnson","year":"2008","journal-title":"Nucleic Acids Res."},{"key":"B25","first-page":"6474","article-title":"Comparison with accurate quantum chemical calculations on peptides \u2020","volume":"2","author":"Kaminski","year":"2001","journal-title":"Quantum"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.01835","article-title":"The human gut microbiome - a potential controller of wellness and disease","volume":"9","author":"Kho","year":"2018","journal-title":"Front. Microbiol."},{"key":"B27","doi-asserted-by":"publisher","first-page":"567","DOI":"10.1006\/jmbi.2000.4315","article-title":"Predicting transmembrane protein topology with a hidden Markov model: Application to complete genomes","volume":"305","author":"Krogh","year":"2001","journal-title":"J. Mol. Biol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"2021.05.02.442313","DOI":"10.1101\/2021.05.02.442313","article-title":"Immunoinformatics approach for a novel multi-epitope vaccine construct against spike protein of human coronaviruses abstract","author":"Kumar","year":"2021","journal-title":"bioRxiv"},{"key":"B29","doi-asserted-by":"publisher","first-page":"1658","DOI":"10.1093\/bioinformatics\/btl158","article-title":"Cd-hit: A fast program for clustering and comparing large sets of protein or nucleotide sequences","volume":"22","author":"Li","year":"2006","journal-title":"Bioinformatics"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1155\/2014\/483905","article-title":"In silico prediction of T and b cell epitopes of der f 25 in dermatophagoides farinae","volume":"2014","author":"Li","year":"2014","journal-title":"Int. J. Genomics"},{"key":"B31","doi-asserted-by":"publisher","first-page":"1153","DOI":"10.1007\/s40265-021-01530-0","article-title":"Epidemiology, treatments, and vaccine development for antimicrobial-resistant neisseria gonorrhoeae: Current strategies and future directions","volume":"81","author":"Lin","year":"2021","journal-title":"Drugs"},{"key":"B32","doi-asserted-by":"publisher","first-page":"D687","DOI":"10.1093\/nar\/gky1080","article-title":"VFDB 2019: A comparative pathogenomic platform with an interactive web interface","volume":"47","author":"Liu","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"B33","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1093\/nar\/gkaa917","article-title":"DEG 15 , an update of the database of essential genes that includes built-in analysis tools","volume":"49","author":"Luo","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"B34","doi-asserted-by":"publisher","first-page":"2200","DOI":"10.1093\/bioinformatics\/btp386","article-title":"SOLpro: accurate sequence-based prediction of protein solubility","volume":"25","author":"Magnan","year":"2009","journal-title":"Bioinformatics"},{"key":"B35","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41598-019-55613-w","article-title":"Designing a multi-epitopic vaccine against the enterotoxigenic bacteroides fragilis based on immunoinformatics approach","volume":"9","author":"Majid","year":"2019","journal-title":"Sci. Rep."},{"key":"B36","doi-asserted-by":"publisher","first-page":"W229","DOI":"10.1093\/nar\/gkn186","article-title":"FireDock: a web server for fast interaction refinement in molecular docking","volume":"36","author":"Mashiach","year":"2008","journal-title":"Nucleic Acids Res."},{"key":"B37","doi-asserted-by":"publisher","first-page":"2459","DOI":"10.1002\/jcc.24904","article-title":"Unfavorable regions in the ramachandran plot: Is it really steric hindrance? the interacting quantum atoms perspective","volume":"38","author":"Maxwell","year":"2017","journal-title":"J. Comput. Chem."},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2020.00054","article-title":"Antibiotic resistance profile, outer membrane proteins, virulence factors and genome sequence analysis reveal clinical isolates of enterobacter are potential pathogens compared to environmental isolates","volume":"10","author":"Mishra","year":"2020","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"B39","doi-asserted-by":"publisher","first-page":"49","DOI":"10.2147\/AABC.S88522","article-title":"Identification of potential drug targets by subtractive genome analysis of escherichia coli O157:H7: An in silico approach","volume":"8","author":"Mondal","year":"2015","journal-title":"Adv. Appl. Bioinforma. Chem."},{"key":"B40","doi-asserted-by":"publisher","first-page":"182","DOI":"10.1093\/nar\/gkm321","article-title":"KAAS: an automatic genome annotation and pathway reconstruction server","volume":"35","author":"Moriya","year":"2007","journal-title":"Nucleic Acids Res."},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0143304","article-title":"Global estimates of the prevalence and incidence of four curable sexually transmitted infections in 2012 based on systematic review and global reporting","volume":"10","author":"Newman","year":"2015","journal-title":"PloS One"},{"key":"B42","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1471-2105-8-238","article-title":"Prediction of MHC class II binding affinity using SMM-align, a novel stabilization matrix alignment method","volume":"8","author":"Nielsen","year":"2007","journal-title":"BMC Bioinf."},{"key":"B43","doi-asserted-by":"publisher","first-page":"2730","DOI":"10.1016\/j.ygeno.2021.06.011","article-title":"Computational identification of putative common genomic drug and vaccine targets in mycoplasma genitalium","volume":"113","author":"Nogueira","year":"2021","journal-title":"Genomics"},{"key":"B44","doi-asserted-by":"publisher","first-page":"135","DOI":"10.15406\/ijmboa.2019.04.00110","article-title":"Bio isolation, chemical purification, identification, antimicrobial and synergistic efficacy of extracted essential oils from stem bark extract of spondias mombin(Linn)","volume":"4","author":"Osuntokun","year":"2019","journal-title":"Int. J. Mol. Biol."},{"key":"B45","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12859-017-1540-0","article-title":"VacSol: A high throughput in silico pipeline to predict potential therapeutic targets in prokaryotic pathogens using subtractive reverse vaccinology","volume":"18","author":"Rizwan","year":"2017","journal-title":"BMC Bioinf."},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1155\/2020\/7672024","article-title":"Prevalence and antimicrobial susceptibility patterns of neisseria gonorrhoeae among suspected patients attending private clinics in jimma, Ethiopia","volume":"2020","author":"Sahile","year":"2020","journal-title":"Int. J. Microbiol."},{"key":"B47","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1155\/2017\/2680160","article-title":"Fundamentals and methods for T- and b-cell epitope prediction","volume":"2017","author":"Sanchez-Trincado","year":"2017","journal-title":"J. Immunol. Res."},{"key":"B48","doi-asserted-by":"publisher","first-page":"181","DOI":"10.1007\/s10989-020-10076-w","article-title":"Delineating novel therapeutic drug and vaccine targets for staphylococcus cornubiensis NW1T through computational analysis","volume":"27","author":"Shah","year":"2021","journal-title":"Int. J. Pept. Res. Ther."},{"key":"B49","doi-asserted-by":"publisher","first-page":"1137","DOI":"10.1177\/1087057114534981","article-title":"Discovery of ATP-competitive inhibitors of tRNAIle lysidine synthetase (TilS) by high-throughput screening","volume":"19","author":"Shapiro","year":"2014","journal-title":"J. Biomol. Screen."},{"key":"B50","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41598-018-26689-7","article-title":"Subtractive proteomics to identify novel drug targets and reverse vaccinology for the development of chimeric vaccine against acinetobacter baumannii","volume":"8","author":"Solanki","year":"2018","journal-title":"Sci. Rep."},{"key":"B51","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/vaccines9060669","article-title":"In silico prediction of a multitope vaccine against moraxella catarrhalis: Reverse vaccinology and immunoinformatics","volume":"9","author":"Soltan","year":"2021","journal-title":"Vaccines"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics10060632","article-title":"In silico prediction and prioritization of novel selective antimicrobial drug targets in escherichia coli","volume":"10","author":"Svanberg Frisinger","year":"2021","journal-title":"Antibiotics"},{"key":"B53","doi-asserted-by":"publisher","first-page":"D362","DOI":"10.1093\/nar\/gkw937","article-title":"The STRING database in 2017: Quality-controlled protein-protein association networks, made broadly accessible","volume":"45","author":"Szklarczyk","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"B54","doi-asserted-by":"publisher","first-page":"11072","DOI":"10.1073\/pnas.1812025115","article-title":"Elongation factor p is required to maintain proteome homeostasis at high growth rate","volume":"115","author":"Tollerson","year":"2018","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B55","doi-asserted-by":"publisher","first-page":"1701","DOI":"10.1002\/jcc.20291","article-title":"GROMACS: Fast, flexible, and free","volume":"26","author":"Van Der Spoel","year":"2005","journal-title":"J. Comput. Chem."},{"key":"B56","doi-asserted-by":"publisher","first-page":"7419","DOI":"10.1016\/j.vaccine.2018.02.081","article-title":"Biological feasibility and importance of a gonorrhea vaccine for global public health","volume":"37","author":"Vincent","year":"2019","journal-title":"Vaccine"},{"key":"B57","doi-asserted-by":"publisher","first-page":"407","DOI":"10.1093\/nar\/gkm290","article-title":"ProSA-web: Interactive web service for the recognition of errors in three-dimensional structures of proteins","volume":"35","author":"Wiederstein","year":"2007","journal-title":"Nucleic Acids Res."},{"key":"B58","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1097\/im9.0000000000000024","article-title":"Antibiotic resistance and treatment options for multidrug-resistant gonorrhea","volume":"2","author":"Yang","year":"2020","journal-title":"Infect. Microbes Dis."},{"key":"B59","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12859-019-3232-4","article-title":"Protein sequence information extraction and subcellular localization prediction with gapped k-mer method","volume":"20","author":"Yao","year":"2019","journal-title":"BMC Bioinf."},{"key":"B60","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s13756-018-0376-3","article-title":"Neisseria gonorrhoae and their antimicrobial susceptibility patterns among symptomatic patients from gondar town, north West Ethiopia","volume":"7","author":"Yeshanew","year":"2018","journal-title":"Antimicrob. Resist. Infect. Control"},{"key":"B61","article-title":"CELLO2GO: a web server for protein subCELlular LOcalization prediction with functional gene ontology annotation","volume":"9","author":"Yu","year":"2014","journal-title":"PLoS One"},{"key":"B62","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1471-2105-9-40","article-title":"I-TASSER server for protein 3D structure prediction","volume":"9","author":"Zhang","year":"2008","journal-title":"BMC Bioinf."},{"key":"B63","doi-asserted-by":"publisher","DOI":"10.3390\/biom11040597","article-title":"Structural basis for the c-terminal domain of mycobacterium tuberculosis ribosome maturation factor rimm to bind ribosomal protein s19","volume":"11","author":"Zhang","year":"2021","journal-title":"Biomolecules"}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1017315\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,3,24]],"date-time":"2023-03-24T05:59:47Z","timestamp":1679637587000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1017315\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,24]]},"references-count":63,"alternative-id":["10.3389\/fcimb.2023.1017315"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2023.1017315","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,24]]},"article-number":"1017315"}}