{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:59:55Z","timestamp":1785419995945,"version":"3.56.0"},"reference-count":40,"publisher":"Springer Science and Business Media LLC","issue":"5","license":[{"start":{"date-parts":[[2022,2,3]],"date-time":"2022-02-03T00:00:00Z","timestamp":1643846400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2022,2,3]],"date-time":"2022-02-03T00:00:00Z","timestamp":1643846400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100005690","name":"Universit\u00e4t des Saarlandes","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100005690","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100005690","name":"Universit\u00e4t des Saarlandes","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100005690","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Mol Biol Rep"],"published-print":{"date-parts":[[2022,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Idiopathic male infertility can be attributed to genetic predispositions that affect sperm performance and function. Genetic alterations in the mitochondrial DNA (mtDNA) have been linked to certain types of male infertility and abnormal sperm function. Mutations in the mitochondrial cytochrome B (<jats:italic>MT-CYB<\/jats:italic>) gene might lead to some deficiencies in mitochondrial function. Thus, in the current study, we aimed to investigate the effect of mutations in the<jats:italic>MT-CYB<\/jats:italic>gene on sperm motility and male infertility.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods and results<\/jats:title><jats:p>Semen specimens were collected from 111 men where 67 men were subfertile and 44 were fertile. QIAamp DNA Mini Kit and REPLI-g Mitochondrial DNA Kit from QIAGEN were used to isolate and amplify the mitochondrial DNA. Followed by PCR and Sanger sequencing for the target sequence in the<jats:italic>MT-CYP<\/jats:italic>gene. Sequencing of the<jats:italic>MT-CYB<\/jats:italic>gene revealed a total of thirteen single nucleotide polymorphisms (SNPs). Eight SNPs were non-synonymous variant (missense variant) including: rs2853508, rs28357685, rs41518645, rs2853507, rs28357376, rs35070048, rs2853506, and rs28660155. While five SNPs were Synonymous variant: rs527236194, rs28357373, rs28357369, rs41504845, and rs2854124. Among these SNPs, three variants showed a significant difference in the frequency of the genotypes between subfertile and fertile groups: rs527236194 (T15784C) (<jats:italic>P<\/jats:italic>\u2009=\u20090.0005), rs28357373 (T15629C) (<jats:italic>P<\/jats:italic>\u2009=\u20090.0439), and rs41504845 (C15833T) (<jats:italic>P<\/jats:italic>\u2009=\u20090.0038). Moreover, two SNPs showed a significant association between allelic frequencies of rs527236194 (T15784C) (<jats:italic>P<\/jats:italic>\u2009=\u20090.0014) and rs41504845 (C15833T) (<jats:italic>P<\/jats:italic>\u2009=\u20090.0147) and male subfertility.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The current study showed a significant association between the<jats:italic>MT-CYB<\/jats:italic>gene polymorphisms and the development of male infertility. In particular, rs527236194, rs28357373 and rs41504845 variants were found to be the most related to the subfertility group. Further studies on larger and other populations are required to reveal the exact role of this gene in the development of male infertility. In addition, functional studies will be helpful to elucidate the molecular impact of the M<jats:italic>T-CYP<\/jats:italic>polymorphisms on mitochondrial function.<\/jats:p><\/jats:sec>","DOI":"10.1007\/s11033-022-07200-y","type":"journal-article","created":{"date-parts":[[2022,2,3]],"date-time":"2022-02-03T22:03:26Z","timestamp":1643925806000},"page":"3609-3616","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Association between the single nucleotide variants of the mitochondrial cytochrome B gene (MT-CYB) and the male infertility"],"prefix":"10.1007","volume":"49","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5847-0465","authenticated-orcid":false,"given":"Mayyas","family":"Saleh Jaweesh","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8513-622X","authenticated-orcid":false,"given":"Mohamad Eid","family":"Hammadeh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2673-2691","authenticated-orcid":false,"given":"Fatina W.","family":"Dahadhah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0248-4777","authenticated-orcid":false,"given":"Mazhar Salim","family":"Al Zoubi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1442-8836","authenticated-orcid":false,"given":"Houda","family":"Amor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2022,2,3]]},"reference":[{"issue":"1","key":"7200_CR1","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1016\/j.fertnstert.2018.03.008","volume":"110","author":"S Fishel","year":"2018","unstructured":"Fishel S (2018) First in vitro fertilization baby\u2014this is how it happened. Fertil Steril 110(1):5\u201311","journal-title":"Fertil Steril"},{"issue":"3","key":"7200_CR2","first-page":"163","volume":"165","author":"D De Berardis","year":"2014","unstructured":"De Berardis D et al (2014) Psychopathology, emotional aspects and psychological counselling in infertility: a review. Clin Ter 165(3):163\u2013169","journal-title":"Clin Ter"},{"key":"7200_CR3","doi-asserted-by":"crossref","unstructured":"Krausz C, Escamilla AR, Chianese C (2015) Genetics of male infertility: from research to clinic. Reproduction 150(5):R159\u2013R174","DOI":"10.1530\/REP-15-0261"},{"key":"7200_CR4","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1038\/nrg1606","volume":"6","author":"RW Taylor","year":"2005","unstructured":"Taylor RW, Turnbull DM (2005) Mitochondrial DNA mutations in human disease. Nat Rev Genet 6:389\u2013402","journal-title":"Nat Rev Genet"},{"issue":"5","key":"7200_CR5","doi-asserted-by":"publisher","first-page":"1540","DOI":"10.1007\/s43032-020-00449-3","volume":"28","author":"MA Al Smadi","year":"2021","unstructured":"Al Smadi MA et al (2021) Impact of mitochondrial genetic variants in ND1, ND2, ND5, and ND6 genes on sperm motility and intracytoplasmic sperm injection (ICSI) outcomes. Reprod Sci 28(5):1540\u20131555","journal-title":"Reprod Sci"},{"issue":"9","key":"7200_CR6","doi-asserted-by":"publisher","first-page":"e13728","DOI":"10.1111\/and.13728","volume":"52","author":"MS Al Zoubi","year":"2020","unstructured":"Al Zoubi MS et al (2020) CAG repeats in the androgen receptor gene is associated with oligozoospermia and teratozoospermia in infertile men in Jordan. Andrologia 52(9):e13728","journal-title":"Andrologia"},{"issue":"2","key":"7200_CR7","doi-asserted-by":"publisher","first-page":"150","DOI":"10.4103\/0970-1591.40606","volume":"24","author":"MB Shamsi","year":"2008","unstructured":"Shamsi MB et al (2008) Mitochondrial DNA mutations in etiopathogenesis of male infertility. Indian J Urol 24(2):150","journal-title":"Indian J Urol"},{"issue":"6","key":"7200_CR8","doi-asserted-by":"publisher","first-page":"1395","DOI":"10.1007\/s00345-013-1023-5","volume":"31","author":"S Shen","year":"2013","unstructured":"Shen S et al (2013) Comparative proteomic study between human normal motility sperm and idiopathic asthenozoospermia. World J Urol 31(6):1395\u20131401","journal-title":"World J Urol"},{"issue":"3","key":"7200_CR9","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1046\/j.1365-2605.2001.00292.x","volume":"24","author":"A Holyoake","year":"2001","unstructured":"Holyoake A et al (2001) High incidence of single nucleotide substitutions in the mitochondrial genome is associated with poor semen parameters in men. Int J Androl 24(3):175\u2013182","journal-title":"Int J Androl"},{"issue":"1","key":"7200_CR10","doi-asserted-by":"crossref","first-page":"5","DOI":"10.4103\/1008-682X.167716","volume":"19","author":"R Pereira","year":"2017","unstructured":"Pereira R et al (2017) Major regulatory mechanisms involved in sperm motility. Asian J Androl 19(1):5","journal-title":"Asian J Androl"},{"issue":"11","key":"7200_CR11","first-page":"GC09","volume":"10","author":"PS Ambulkar","year":"2016","unstructured":"Ambulkar PS et al (2016) Large scale 7436-bp deletions in human sperm mitochondrial DNA with spermatozoa dysfunction and male infertility. J Clin Diagn Res 10(11):GC09","journal-title":"J Clin Diagn Res"},{"issue":"1","key":"7200_CR12","doi-asserted-by":"publisher","first-page":"e13379","DOI":"10.1111\/and.13379","volume":"52","author":"MS Al Zoubi","year":"2020","unstructured":"Al Zoubi MS et al (2020) 4,977-bp human mitochondrial DNA deletion is associated with asthenozoospermic infertility in Jordan. Andrologia 52(1):e13379","journal-title":"Andrologia"},{"issue":"7","key":"7200_CR13","doi-asserted-by":"publisher","first-page":"2315","DOI":"10.1016\/j.fertnstert.2011.03.059","volume":"95","author":"D Paoli","year":"2011","unstructured":"Paoli D et al (2011) Mitochondrial membrane potential profile and its correlation with increasing sperm motility. Fertil Steril 95(7):2315\u20132319","journal-title":"Fertil Steril"},{"issue":"9","key":"7200_CR14","doi-asserted-by":"publisher","first-page":"1043","DOI":"10.1161\/CIRCRESAHA.113.301151","volume":"113","author":"NC Zong","year":"2013","unstructured":"Zong NC et al (2013) Integration of cardiac proteome biology and medicine by a specialized knowledgebase. Circ Res 113(9):1043\u20131053","journal-title":"Circ Res"},{"issue":"2","key":"7200_CR15","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1016\/j.bbabio.2009.09.005","volume":"1797","author":"HA Tuppen","year":"2010","unstructured":"Tuppen HA et al (2010) Mitochondrial DNA mutations and human disease. Biochim Biophys Acta 1797(2):113\u2013128","journal-title":"Biochim Biophys Acta"},{"issue":"4","key":"7200_CR16","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1136\/jmg.2003.015289","volume":"41","author":"M Giacchetti","year":"2004","unstructured":"Giacchetti M et al (2004) Mitochondrial DNA haplogroups influence the Friedreich\u2019s ataxia phenotype. J Med Genet 41(4):293\u2013295","journal-title":"J Med Genet"},{"issue":"2","key":"7200_CR17","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1038\/13779","volume":"23","author":"RM Andrews","year":"1999","unstructured":"Andrews RM et al (1999) Reanalysis and revision of the Cambridge reference sequence for human mitochondrial DNA. Nat Genet 23(2):147\u2013147","journal-title":"Nat Genet"},{"key":"7200_CR18","unstructured":"Kumar R, Venkatesh S, Kumar M, Tanwar M, Shasmsi MB, Gupta NP, Sharma RK, Talwar P, Dada R (2009) Oxidative stress and sperm mitochondrial DNA mutation in idiopathic oligoasthenozoospermic men"},{"issue":"2","key":"7200_CR19","first-page":"127","volume":"129","author":"S Venkatesh","year":"2009","unstructured":"Venkatesh S et al (2009) Role of reactive oxygen species in the pathogenesis of mitochondrial DNA (mtDNA) mutations in male infertility. Indian J Med Res 129(2):127\u2013137","journal-title":"Indian J Med Res"},{"issue":"10","key":"7200_CR20","doi-asserted-by":"publisher","first-page":"1876","DOI":"10.1093\/humrep\/dez153","volume":"34","author":"M Xavier","year":"2019","unstructured":"Xavier M et al (2019) Paternal impacts on development: identification of genomic regions vulnerable to oxidative DNA damage in human spermatozoa. Hum Reprod 34(10):1876\u20131890","journal-title":"Hum Reprod"},{"issue":"3","key":"7200_CR21","doi-asserted-by":"publisher","first-page":"897","DOI":"10.1093\/genetics\/129.3.897","volume":"129","author":"M Bulmer","year":"1991","unstructured":"Bulmer M (1991) The selection-mutation-drift theory of synonymous codon usage. Genetics 129(3):897\u2013907","journal-title":"Genetics"},{"key":"7200_CR22","first-page":"2687","volume":"18","author":"M Kumar","year":"2012","unstructured":"Kumar M et al (2012) Clinical characterization and mitochondrial DNA sequence variations in Leber hereditary optic neuropathy. Mol Vis 18:2687","journal-title":"Mol Vis"},{"issue":"1","key":"7200_CR23","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41598-019-45934-1","volume":"9","author":"E Fasterius","year":"2019","unstructured":"Fasterius E, Uhl\u00e9n M, Szigyarto CA-K (2019) Single-cell RNA-seq variant analysis for exploration of genetic heterogeneity in cancer. Sci Rep 9(1):1\u201311","journal-title":"Sci Rep"},{"issue":"4","key":"7200_CR24","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1007\/BF01247339","volume":"92","author":"K Huoponen","year":"1993","unstructured":"Huoponen K et al (1993) The spectrum of mitochondrial DNA mutations in families with Leber hereditary optic neuroretinopathy. Hum Genet 92(4):379\u2013384","journal-title":"Hum Genet"},{"issue":"9","key":"7200_CR25","doi-asserted-by":"publisher","first-page":"e0136192","DOI":"10.1371\/journal.pone.0136192","volume":"10","author":"S Blein","year":"2015","unstructured":"Blein S et al (2015) Targeted sequencing of the mitochondrial genome of women at high risk of breast cancer without detectable mutations in BRCA1\/2. PLoS ONE 10(9):e0136192","journal-title":"PLoS ONE"},{"issue":"4","key":"7200_CR26","first-page":"321","volume":"14","author":"C-Q Feng","year":"2008","unstructured":"Feng C-Q et al (2008) Mutation of MTCYB and MTATP6 is associated with asthenospermia. Natl J Androl 14(4):321\u2013323","journal-title":"Natl J Androl"},{"issue":"1","key":"7200_CR27","doi-asserted-by":"publisher","first-page":"20","DOI":"10.3109\/19401736.2013.840612","volume":"26","author":"G-H Mao","year":"2015","unstructured":"Mao G-H et al (2015) Polymorphisms in the MT-ATP6 and MT-CYB genes in in vitro fertilization failure. Mitochondrial DNA 26(1):20\u201324","journal-title":"Mitochondrial DNA"},{"issue":"1","key":"7200_CR28","first-page":"67","volume":"25","author":"MLM Rezgoune","year":"2021","unstructured":"Rezgoune MLM et al (2021) Association of the CAG repeat polymorphism in mitochondrial polymerase gamma (POLG1) with male infertility: a case-control study in an Algerian population. Afr J Reprod Health 25(1):67\u201375","journal-title":"Afr J Reprod Health"},{"issue":"1","key":"7200_CR29","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1471-2350-12-50","volume":"12","author":"V \u00c1lvarez-Iglesias","year":"2011","unstructured":"\u00c1lvarez-Iglesias V et al (2011) Reassessing the role of mitochondrial DNA mutations in autism spectrum disorder. BMC Med Genet 12(1):1\u20137","journal-title":"BMC Med Genet"},{"issue":"1","key":"7200_CR30","first-page":"265","volume":"4","author":"PN Barbhuiya","year":"2016","unstructured":"Barbhuiya PN et al (2016) Prevalence of mitochondrial DNA nucleotide substitution mutations in male infertile cases of northeast India. J Infertil Reprod Biol 4(1):265\u2013275","journal-title":"J Infertil Reprod Biol"},{"issue":"3","key":"7200_CR31","doi-asserted-by":"publisher","first-page":"738","DOI":"10.1016\/j.fertnstert.2007.03.048","volume":"89","author":"L Pereira","year":"2008","unstructured":"Pereira L, Gon\u00e7alves J, Bandelt H-J (2008) Mutation C11994T in the mitochondrial ND4 gene is not a cause of low sperm motility in Portugal. Fertil Steril 89(3):738\u2013741","journal-title":"Fertil Steril"},{"issue":"1","key":"7200_CR32","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s43141-020-00111-0","volume":"19","author":"MM Abd Elrahman","year":"2021","unstructured":"Abd Elrahman MM et al (2021) Assessment of correlation between asthenozoospermia and mitochondrial DNA mutations in Egyptian infertile men. J Genet Eng Biotechnol 19(1):1\u201315","journal-title":"J Genet Eng Biotechnol"},{"key":"7200_CR33","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s10815-021-02199-w","volume":"38","author":"FW Dahadhah","year":"2021","unstructured":"Dahadhah FW et al (2021) Mitochondrial nicotinamide adenine dinucleotide hydride dehydrogenase (NADH) subunit 4 (MTND4) polymorphisms and their association with male infertility. J Assist Reprod Genet 38:1\u20139","journal-title":"J Assist Reprod Genet"},{"issue":"3","key":"7200_CR34","doi-asserted-by":"publisher","first-page":"928","DOI":"10.3906\/sag-1606-52","volume":"47","author":"IA Mughal","year":"2017","unstructured":"Mughal IA et al (2017) Male infertility is significantly associated with multiple deletions in an 8.7-kb segment of sperm mtDNA in Pakistan. Turk J Med Sci 47(3):928\u2013933","journal-title":"Turk J Med Sci"},{"key":"7200_CR35","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1016\/j.ebiom.2015.11.045","volume":"3","author":"AUH Khan","year":"2016","unstructured":"Khan AUH et al (2016) Human leukemic cells performing oxidative phosphorylation (OXPHOS) generate an antioxidant response independently of reactive oxygen species (ROS) production. EBioMedicine 3:43\u201353","journal-title":"EBioMedicine"},{"issue":"6","key":"7200_CR36","doi-asserted-by":"publisher","first-page":"665","DOI":"10.1007\/s00439-017-1779-6","volume":"136","author":"PD Stenson","year":"2017","unstructured":"Stenson PD et al (2017) The human gene mutation database: towards a comprehensive repository of inherited mutation data for medical research, genetic diagnosis and next-generation sequencing studies. Hum Genet 136(6):665\u2013677","journal-title":"Hum Genet"},{"issue":"5","key":"7200_CR37","doi-asserted-by":"publisher","first-page":"725","DOI":"10.1080\/24701394.2016.1177038","volume":"28","author":"Y Zhang","year":"2017","unstructured":"Zhang Y et al (2017) Associations of mitochondrial haplogroups and mitochondrial DNA copy numbers with end-stage renal disease in a Han population. Mitochondrial DNA A 28(5):725\u2013731","journal-title":"Mitochondrial DNA A"},{"issue":"1","key":"7200_CR38","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1212\/WNL.49.1.239","volume":"49","author":"R Fadic","year":"1997","unstructured":"Fadic R et al (1997) Sensory ataxic neuropathy as the presenting feature of a novel mitochondrial disease. Neurology 49(1):239\u2013245","journal-title":"Neurology"},{"issue":"9","key":"7200_CR39","doi-asserted-by":"publisher","first-page":"811","DOI":"10.1093\/molehr\/3.9.811","volume":"3","author":"P Lestienne","year":"1997","unstructured":"Lestienne P et al (1997) Oligoasthenospermia associated with multiple mitochondrial DNA rearrangements. Mol Hum Reprod 3(9):811\u2013814","journal-title":"Mol Hum Reprod"},{"issue":"7","key":"7200_CR40","doi-asserted-by":"publisher","first-page":"657","DOI":"10.1093\/molehr\/4.7.657","volume":"4","author":"S-H Kao","year":"1998","unstructured":"Kao S-H, Chao H-T, Wei Y-H (1998) Multiple deletions of mitochondrial DNA are associated with the decline of motility and fertility of human spermatozoa. Mol Hum Reprod 4(7):657\u2013666","journal-title":"Mol Hum Reprod"}],"container-title":["Molecular Biology Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11033-022-07200-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s11033-022-07200-y\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11033-022-07200-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,16]],"date-time":"2023-11-16T22:43:51Z","timestamp":1700174631000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s11033-022-07200-y"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,3]]},"references-count":40,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2022,5]]}},"alternative-id":["7200"],"URL":"https:\/\/doi.org\/10.1007\/s11033-022-07200-y","relation":{},"ISSN":["0301-4851","1573-4978"],"issn-type":[{"value":"0301-4851","type":"print"},{"value":"1573-4978","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,3]]},"assertion":[{"value":"8 October 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 January 2022","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 February 2022","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"All authors declare there is no conflict of interest in this work.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and\/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The name of the committee that ethically approved this study is: Jordanian Royal Medical Services-Human Research Ethics Committee The Approval Number is F3\/1\/ Ethics Committee \/ 9126.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}},{"value":"Informed consent was obtained from all individual participants included in the study.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Informed consent"}}]}}