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Article ; Online: Sperm DNA damage measured by the alkaline Comet assay as an independent predictor of male infertility and in vitro fertilization success.

Simon, Luke / Lutton, Deborah / McManus, Joanne / Lewis, Sheena E M

2011 Volume 95, Issue 2, Page(s) 652–657

Abstract: Objective: To evaluate sperm DNA fragmentation and semen parameters to diagnose male factor infertility and predict pregnancy after IVF.: Design: Prospective study.: Setting: Academic research laboratory.: Patient(s): Seventy-five couples ... ...

Abstract	Objective: To evaluate sperm DNA fragmentation and semen parameters to diagnose male factor infertility and predict pregnancy after IVF. Design: Prospective study. Setting: Academic research laboratory. Patient(s): Seventy-five couples undergoing IVF and 28 fertile donors. Intervention(s): Sperm DNA fragmentation was measured by the alkaline Comet assay in semen and sperm after density gradient centrifugation (DGC). Binary logistic regression was used to analyze odds ratios (OR) and relative risks (RR) for IVF outcomes. Main outcome measure(s): Semen parameters and sperm DNA fragmentation in semen and DGC sperm compared with fertilization rates, embryo quality, and pregnancy. Result(s): Men with sperm DNA fragmentation at more than a diagnostic threshold of 25% had a high risk of infertility (OR: 117.33, 95% confidence interval [CI]: 12.72-2,731.84, RR: 8.75). Fertilization rates and embryo quality decreased as sperm DNA fragmentation increased in semen and DGC sperm. The risk of failure to achieve a pregnancy increased when sperm DNA fragmentation exceeded a prognostic threshold value of 52% for semen (OR: 76.00, CI: 8.69-1,714.44, RR: 4.75) and 42% for DGC sperm (OR: 24.18, CI: 2.89-522.34, RR: 2.16). Conclusion(s): Sperm DNA testing by the alkaline Comet assay is useful for both diagnosis of male factor infertility and prediction of IVF outcome.
MeSH term(s)	Adult ; Comet Assay/methods ; DNA Damage/physiology ; Female ; Fertilization in Vitro/methods ; Humans ; Infertility, Male/diagnosis ; Infertility, Male/genetics ; Infertility, Male/pathology ; Infertility, Male/therapy ; Male ; Middle Aged ; Pregnancy ; Prognosis ; Semen Analysis/methods ; Spermatozoa/metabolism ; Spermatozoa/pathology ; Treatment Outcome ; Young Adult
Language	English
Publishing date	2011-02
Publishing country	United States
Document type	Clinical Trial ; Journal Article
ZDB-ID	80133-1
ISSN	1556-5653 ; 0015-0282
ISSN (online)	1556-5653
ISSN	0015-0282
DOI	10.1016/j.fertnstert.2010.08.019
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Article: Sperm DNA damage measured by the alkaline Comet assay as an independent predictor of male infertility and in vitro fertilization success

Simon, Luke / Lutton, Deborah / McManus, Joanne / Lewis, Sheena E.M

Fertility and sterility. 2011 Feb., v. 95, no. 2

2011

Abstract: OBJECTIVE: To evaluate sperm DNA fragmentation and semen parameters to diagnose male factor infertility and predict pregnancy after IVF. DESIGN: Prospective study. SETTING: Academic research laboratory. PATIENT(S): Seventy-five couples undergoing IVF and ...

Abstract	OBJECTIVE: To evaluate sperm DNA fragmentation and semen parameters to diagnose male factor infertility and predict pregnancy after IVF. DESIGN: Prospective study. SETTING: Academic research laboratory. PATIENT(S): Seventy-five couples undergoing IVF and 28 fertile donors. INTERVENTION(S): Sperm DNA fragmentation was measured by the alkaline Comet assay in semen and sperm after density gradient centrifugation (DGC). Binary logistic regression was used to analyze odds ratios (OR) and relative risks (RR) for IVF outcomes. MAIN OUTCOME MEASURE(S): Semen parameters and sperm DNA fragmentation in semen and DGC sperm compared with fertilization rates, embryo quality, and pregnancy. RESULT(S): Men with sperm DNA fragmentation at more than a diagnostic threshold of 25% had a high risk of infertility (OR: 117.33, 95% confidence interval [CI]: 12.72–2,731.84, RR: 8.75). Fertilization rates and embryo quality decreased as sperm DNA fragmentation increased in semen and DGC sperm. The risk of failure to achieve a pregnancy increased when sperm DNA fragmentation exceeded a prognostic threshold value of 52% for semen (OR: 76.00, CI: 8.69–1,714.44, RR: 4.75) and 42% for DGC sperm (OR: 24.18, CI: 2.89–522.34, RR: 2.16). CONCLUSION(S): Sperm DNA testing by the alkaline Comet assay is useful for both diagnosis of male factor infertility and prediction of IVF outcome.
Keywords	DNA ; DNA damage ; DNA fragmentation ; confidence interval ; density gradient centrifugation ; in vitro fertilization ; male fertility ; men ; odds ratio ; patients ; prediction ; pregnancy ; prospective studies ; regression analysis ; risk ; semen ; spermatozoa
Language	English
Dates of publication	2011-02
Size	p. 652-657.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	80133-1
ISSN	1556-5653 ; 0015-0282
ISSN (online)	1556-5653
ISSN	0015-0282
DOI	10.1016/j.fertnstert.2010.08.019
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Clinical significance of sperm DNA damage in assisted reproduction outcome.

Simon, Luke / Brunborg, Gunnar / Stevenson, Michael / Lutton, Deborah / McManus, Joanne / Lewis, Sheena E M

Human reproduction (Oxford, England)

2010 Volume 25, Issue 7, Page(s) 1594–1608

Abstract: Background: Sperm DNA damage shows great promise as a biomarker of infertility. The study aim is to determine the usefulness of DNA fragmentation (DF), including modified bases (MB), to predict assisted reproduction treatment (ART) outcomes.: Methods!# ...

Abstract	Background: Sperm DNA damage shows great promise as a biomarker of infertility. The study aim is to determine the usefulness of DNA fragmentation (DF), including modified bases (MB), to predict assisted reproduction treatment (ART) outcomes. Methods: DF in 360 couples (230 IVF and 130 ICSI) was measured by the alkaline Comet assay in semen and in sperm following density gradient centrifugation (DGC) and compared with fertilization rate (FR), embryo cumulative scores (ECS(1)) for the total number of embryos/treatment, embryos transferred (ECS(2)), clinical pregnancy (CP) and spontaneous pregnancy loss. MB were also measured using formamidopyrimidine DNA glycosylase to convert them into strand breaks. Results: In IVF, FR and ECS decreased as DF increased in both semen and DGC sperm, and couples who failed to achieve a CP had higher DF than successful couples (+12.2% semen, P = 0.004; +9.9% DGC sperm, P = 0.010). When MB were added to existing strand breaks, total DF was markedly higher (+17.1% semen, P = 0.009 and +13.8% DGC sperm, P = 0.045). DF was not associated with FR, ECS or CP in either semen or DGC sperm following ISCI. In contrast, by including MB, there was significantly more DNA damage (+16.8% semen, P = 0.008 and +15.5% DGC sperm, P = 0.024) in the group who did not achieve CP. Conclusions: DF can predict ART outcome for IVF. Converting MB into further DNA strand breaks increased the test sensitivity, giving negative correlations between DF and CP for ICSI as well as IVF.
MeSH term(s)	Abortion, Spontaneous ; Comet Assay ; DNA Fragmentation ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility/therapy ; Male ; Predictive Value of Tests ; Pregnancy ; Pregnancy Rate ; Semen Analysis ; Spermatozoa/physiology ; Treatment Outcome
Language	English
Publishing date	2010-07
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	632776-x
ISSN	1460-2350 ; 0268-1161 ; 1477-741X
ISSN (online)	1460-2350
ISSN	0268-1161 ; 1477-741X
DOI	10.1093/humrep/deq103
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Differences in the endocannabinoid system of sperm from fertile and infertile men.

Lewis, Sheena E M / Rapino, Cinzia / Di Tommaso, Monia / Pucci, Mariangela / Battista, Natalia / Paro, Rita / Simon, Luke / Lutton, Deborah / Maccarrone, Mauro

PloS one

2012 Volume 7, Issue 10, Page(s) e47704

Abstract: Male infertility is a major cause of problems for many couples in conceiving a child. Recently, lifestyle pastimes such as alcohol, tobacco and marijuana have been shown to have further negative effects on male reproduction. The endocannabinoid system ( ... ...

Abstract	Male infertility is a major cause of problems for many couples in conceiving a child. Recently, lifestyle pastimes such as alcohol, tobacco and marijuana have been shown to have further negative effects on male reproduction. The endocannabinoid system (ECS), mainly through the action of anandamide (AEA) and 2-arachidonoylglycerol (2-AG) at cannabinoid (CB(1), CB(2)) and vanilloid (TRPV1) receptors, plays a crucial role in controlling functionality of sperm, with a clear impact on male reproductive potential. Here, sperm from fertile and infertile men were used to investigate content (through LC-ESI-MS), mRNA (through quantitative RT-PCR), protein (through Western Blotting and ELISA) expression, and functionality (through activity and binding assays) of the main metabolic enzymes of AEA and 2-AG (NAPE-PLD and FAAH, for AEA; DAGL and MAGL for 2-AG), as well as of their binding receptors CB(1), CB(2) and TRPV1. Our findings show a marked reduction of AEA and 2-AG content in infertile seminal plasma, paralleled by increased degradation: biosynthesis ratios of both substances in sperm from infertile versus fertile men. In addition, TRPV1 binding was detected in fertile sperm but was undetectable in infertile sperm, whereas that of CB(1) and CB(2) receptors was not statistically different in the two groups. In conclusion, this study identified unprecedented alterations of the ECS in infertile sperm, that might impact on capacitation and acrosome reaction, and hence fertilization outcomes. These alterations might also point to new biomarkers to determine male reproductive defects, and identify distinct ECS elements as novel targets for therapeutic exploitation of ECS-oriented drugs to treat male fertility problems.
MeSH term(s)	Adult ; Demography ; Endocannabinoids/genetics ; Endocannabinoids/metabolism ; Fertility/genetics ; Gene Expression Regulation ; Humans ; Infertility, Male/genetics ; Infertility, Male/metabolism ; Male ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Semen/metabolism ; Spermatozoa/metabolism ; Spermatozoa/pathology
Chemical Substances	Endocannabinoids ; RNA, Messenger
Language	English
Publishing date	2012-10-17
Publishing country	United States
Document type	Journal Article
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0047704
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Effects of short and long incubations on DNA fragmentation of testicular sperm.

Dalzell, Lauren H / McVicar, Carmel M / McClure, Neil / Lutton, Deborah / Lewis, Sheena E M

Fertility and sterility

2004 Volume 82, Issue 5, Page(s) 1443–1445

Abstract: DNA fragmentation in testicular sperm from men with obstructive azoospermia is increased by 4-hour and 24-hour incubations and after cryopreservation with the effect is intensified by post-thaw incubation. Testicular sperm for use in intracytoplasmic ... ...

Abstract	DNA fragmentation in testicular sperm from men with obstructive azoospermia is increased by 4-hour and 24-hour incubations and after cryopreservation with the effect is intensified by post-thaw incubation. Testicular sperm for use in intracytoplasmic sperm injection (ICSI) should be injected without delay.
MeSH term(s)	Cell Culture Techniques ; Cryopreservation ; DNA Fragmentation ; Humans ; Male ; Oligospermia/genetics ; Spermatozoa ; Testis ; Time Factors
Language	English
Publishing date	2004-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	80133-1
ISSN	1556-5653 ; 0015-0282
ISSN (online)	1556-5653
ISSN	0015-0282
DOI	10.1016/j.fertnstert.2004.04.053
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Article: Immune reactions to Bacteroides fragilis populations with three different types of capsule in a model of infection.

Patrick, Sheila / Lutton, Deborah A / Crockard, Alistair D

Microbiology (Reading, England)

1995 Volume 141 Pt 8, Page(s) 1969–1976

Abstract: The survival and growth of populations of the obligately anaerobic pathogenic bacterium Bacteroides fragilis enriched for large capsules (LCs), small capsules (SCs) or an electron-dense layer (EDL; non-capsulate by light microscopy) were examined in a ... ...

Abstract	The survival and growth of populations of the obligately anaerobic pathogenic bacterium Bacteroides fragilis enriched for large capsules (LCs), small capsules (SCs) or an electron-dense layer (EDL; non-capsulate by light microscopy) were examined in a mouse model of infection over a minimum period of 20 d. Chambers which allowed the influx of leukocytes, but not the efflux of bacteria, were implanted in the mouse peritoneal cavity. The LC and EDL populations consistently attained viable cell densities of the order of 10(8)-10(9) c.f.u. ml-1 within 24 h, whereas the SC population did not. However, after 3 d, all three bacterial populations maintained total viable numbers of 10(8)-10(9) c.f.u. ml-1 within the chambers. LC expression was selected against within 24 h in the model, the populations becoming non-capsulate by light microscopy, whereas in the SC population expression of the SC was retained by approximately 90% of the population. The EDL population remained non-capsulate by light microscopy throughout. Lymphocytes infiltrated the chambers to an equal extent for all three B. fragilis populations and at approximately 1000 times higher concentration than chambers which contained only quarter-strength Ringer's solution. The presence of neutrophils within the chambers did not cause a decrease in the total viable bacterial count. Each population elicited antibodies specific for outer-membrane proteins and polysaccharide, as detected by immunoblotting, which cross-reacted with the other populations. Differences were observed in the immunogenicity of the outer-membrane proteins within the three populations. Neutrophils were initially the predominant cell type in the chambers, but as the total leukocyte count increased with incubation time, neutrophils were outnumbered by other leukocytes.(ABSTRACT TRUNCATED AT 250 WORDS)
MeSH term(s)	Animals ; Antibodies, Bacterial/biosynthesis ; B-Lymphocytes/immunology ; Bacterial Capsules/chemistry ; Bacterial Capsules/immunology ; Bacterial Outer Membrane Proteins/immunology ; Bacteroidaceae Infections/immunology ; Bacteroides fragilis/genetics ; Bacteroides fragilis/growth & development ; Bacteroides fragilis/immunology ; Bacteroides fragilis/metabolism ; Epitopes/immunology ; Immune Sera/immunology ; Immunity, Cellular ; Immunoblotting ; Leukocyte Count ; Mice ; Neutrophils/immunology ; Polysaccharides, Bacterial/immunology ; Time Factors
Chemical Substances	Antibodies, Bacterial ; Bacterial Outer Membrane Proteins ; Epitopes ; Immune Sera ; Polysaccharides, Bacterial
Language	English
Publishing date	1995-08
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1180712-x
ISSN	1465-2080 ; 1350-0872
ISSN (online)	1465-2080
ISSN	1350-0872
DOI	10.1099/13500872-141-8-1969
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Article: Sperm DNA damage measured by the alkaline Comet assay as an independent predictor of male infertility and in vitro fertilization success

Simon, Luke / Lutton, Deborah / McManus, Joanne / Lewis, Sheena E.M.

Fertility and sterility

Volume v. 95,, Issue no. 2

Abstract	OBJECTIVE: To evaluate sperm DNA fragmentation and semen parameters to diagnose male factor infertility and predict pregnancy after IVF. DESIGN: Prospective study. SETTING: Academic research laboratory. PATIENT(S): Seventy-five couples undergoing IVF and 28 fertile donors. INTERVENTION(S): Sperm DNA fragmentation was measured by the alkaline Comet assay in semen and sperm after density gradient centrifugation (DGC). Binary logistic regression was used to analyze odds ratios (OR) and relative risks (RR) for IVF outcomes. MAIN OUTCOME MEASURE(S): Semen parameters and sperm DNA fragmentation in semen and DGC sperm compared with fertilization rates, embryo quality, and pregnancy. RESULT(S): Men with sperm DNA fragmentation at more than a diagnostic threshold of 25% had a high risk of infertility (OR: 117.33, 95% confidence interval [CI]: 12.72–2,731.84, RR: 8.75). Fertilization rates and embryo quality decreased as sperm DNA fragmentation increased in semen and DGC sperm. The risk of failure to achieve a pregnancy increased when sperm DNA fragmentation exceeded a prognostic threshold value of 52% for semen (OR: 76.00, CI: 8.69–1,714.44, RR: 4.75) and 42% for DGC sperm (OR: 24.18, CI: 2.89–522.34, RR: 2.16). CONCLUSION(S): Sperm DNA testing by the alkaline Comet assay is useful for both diagnosis of male factor infertility and prediction of IVF outcome.
Keywords	prospective studies ; semen ; confidence interval ; prediction ; male fertility ; risk ; DNA damage ; DNA fragmentation ; patients ; spermatozoa ; in vitro fertilization ; odds ratio ; pregnancy ; DNA ; density gradient centrifugation ; men ; regression analysis
Language	English
Document type	Article
ISSN	0015-0282
Database	AGRIS - International Information System for the Agricultural Sciences and Technology

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Article ; Online: Sperm DNA damage measured by the alkaline Comet assay as an independent predictor of male infertility and in vitro fertilization success.

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Article ; Online: Clinical significance of sperm DNA damage in assisted reproduction outcome.

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Article ; Online: Differences in the endocannabinoid system of sperm from fertile and infertile men.

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Article: Effects of short and long incubations on DNA fragmentation of testicular sperm.

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Article: Immune reactions to Bacteroides fragilis populations with three different types of capsule in a model of infection.

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Article: Sperm DNA damage measured by the alkaline Comet assay as an independent predictor of male infertility and in vitro fertilization success

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