title: A simple sperm DNA toroid integrity test and risk of miscarriage.
abstract: Current methods of analyzing sperm chromatin competency overlook the inner sperm compartment which is inaccessible to probes and reagents. By breaking the molecular protamine disulfide bridges, the DNA toroids are exposed to integrity analysis. The aim was to develop a simple nuclear toroid test and determine its association with fertilization, pregnancy, and miscarriage. The approach involved treating washed sperm remaining after ICSI procedures (N=35 cases) with acidified Triton X-100 and dithiothreitol (DTT) before Diff-Quik staining. Percentages of sperm with normal chromatin indicated by light-colored nuclei were assessed. The toroid integrity test showed more sperm with normal chromatin in the pregnant group (73.6±1.7%, mean±SEM) when compared with the miscarriage (51.2±6.6%) or nonpregnant groups (60.9±4.8%). Furthermore, the toroid results were correlated with ICSI fertilization (R=0.32, P=0.04) and pregnancy outcome (pregnant cases 73.6±1.7% versus nonpregnant 58.0±3.9%, P=0.001). ROC calculated cut-off was >70.0% for normal toroid integrity (sensitivity 0.98, specificity 0.33, and diagnostic accuracy 78.3%). An association between normal sperm toroid integrity and miscarriage was evident when the staining procedure included acidified detergent DTT pretreatment.
authors: Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, Loma Linda, CA 92354, USA.
authors: Philip J
authors: PJ
authors: Chan
authors: Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, Loma Linda, CA 92354, USA.
authors: Eliza M
authors: EM
authors: Orzylowska
authors: Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, Loma Linda, CA 92354, USA.
authors: Johannah U
authors: JU
authors: Corselli
authors: Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, Loma Linda, CA 92354, USA.
authors: John D
authors: JD
authors: Jacobson
authors: Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, Loma Linda, CA 92354, USA.
authors: Albert K
authors: AK
authors: Wei
mesh_terms: D000022:Abortion, Spontaneous / Q000453:epidemiology*; D000328:Adult; D002467:Cell Nucleus / Q000737:chemistry; D002843:Chromatin / Q000737:chemistry / Q000235:genetics; D004247:DNA / Q000032:analysis / Q000737:chemistry; D005260:Female; D006801:Humans; D008297:Male; D011247:Pregnancy; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000737:chemistry / Q000166:cytology / Q000473:pathology
chemical_list: D002843:Chromatin; D004247:DNA
references: Mol Reprod Dev. 2000 Jun;56(2 Suppl):230-4
references: 10824973
references: Fertil Steril. 2008 Aug;90(2):352-9
references: 17904130
references: Hum Reprod. 2008 Dec;23(12):2663-8
references: 18757447
references: Hum Reprod. 2009 Jan;24(1):28-36
references: 18835873
references: Mol Hum Reprod. 2010 Jan;16(1):30-6
references: 19748904
references: Int J Androl. 2011 Feb;34(1):2-13
references: 20158539
references: Fertil Steril. 2011 Feb;95(2):652-7
references: 20864101
references: Nat Cell Biol. 2011 Oct;13(10):1161-9
references: 21968989
references: Andrologia. 2012 May;44 Suppl 1:462-70
references: 21806662
references: Biophys J. 2012 Jul 3;103(1):118-28
references: 22828338
references: Hum Reprod. 2012 Oct;27(10):2908-17
references: 22791753
references: Mol Hum Reprod. 2013 Aug;19(8):475-85
references: 23548339
references: Reprod Biomed Online. 2013 Oct;27(4):325-37
references: 23948450
references: Biochim Biophys Acta. 2014 Mar;1839(3):155-68
references: 24091090
references: Mol Hum Reprod. 2014 Apr;20(4):330-40
references: 24282283
references: Hum Reprod Update. 2003 Jul-Aug;9(4):331-45
references: 12926527
references: Hum Reprod. 2000 Jun;15(6):1338-44
references: 10831565
references: J Androl. 2000 Nov-Dec;21(6):903-12
references: 11105917
references: Arch Androl. 2000 Nov-Dec;45(3):215-25
references: 11111870
references: Biol Reprod. 2002 Nov;67(5):1532-7
references: 12390885
references: J Androl. 2003 Jan-Feb;24(1):59-66
references: 12514084
references: Fertil Steril. 2004 May;81(5):1289-95
references: 15136092
references: Reprod Fertil Dev. 2004;16(5):547-54
references: 15355763
references: Andrologia. 2004 Oct;36(5):321-6
references: 15458552
references: Fertil Steril. 1984 Jul;42(1):87-91
references: 6724015
references: Urology. 1987 Sep;30(3):248-51
references: 3629768
references: Exp Cell Res. 1988 Mar;175(1):184-91
references: 3345800
references: Biol Reprod. 1991 Apr;44(4):569-74
references: 2043729
references: Biol Reprod. 1993 Jun;48(6):1193-201
references: 8318576
references: Acta Histochem. 1993 May;94(2):141-9
references: 7688924
references: Methods Cell Biol. 1994;42 Pt B:159-76
references: 7533238
references: Int J Androl. 1995 Jun;18 Suppl 1:23-8
references: 7558384
references: Biol Reprod. 1997 Jan;56(1):42-9
references: 9002631
references: J Assist Reprod Genet. 1997 Aug;14(7):388-93
references: 9285323
references: Hum Reprod Update. 1997 Jul-Aug;3(4):393-400
references: 9459284
references: Int J Androl. 1998 Feb;21(1):13-8
references: 9639147
references: Biol Reprod. 1999 Mar;60(3):702-6
references: 10026119
references: Science. 1999 Oct 1;286(5437):120-3
references: 10506559
references: Asian J Androl. 2006 Jan;8(1):11-29
references: 16372115
references: J Androl. 2006 Jan-Feb;27(1):53-9
references: 16400078
references: CMAJ. 2006 Aug 29;175(5):495-500
references: 16940270
references: Hum Reprod. 2007 Jan;22(1):174-9
references: 16921163
references: Fertil Steril. 2007 Jan;87(1):93-100
references: 17074327
references: Hum Reprod. 2007 May;22(5):1431-42
references: 17303633
references: J Assist Reprod Genet. 2007 Jul;24(7):288-95
references: 17629722
references: Fertil Steril. 2008 Apr;89(4):823-31
references: 17644094
references: Syst Biol Reprod Med. 2008 May-Jun;54(3):111-25
references: 18570047
references: Fertil Steril. 2008 Nov;90(5):1792-9
references: 18166175