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The relationship between DNA fragmentation and the intensity of morphologically abnormal human spermatozoa

Asian pacific Journal of Reproduction 2024年第1期13卷 22-27页

作者： Mercedes González-Martínez Pascual Sánchez-Martín Carmen López-Fernández Stephen D.Johnston Jaime Gosálvez Clínica Ginemed Unidad de ReproducciónSevillaSpain Department of Biology Universidad Autónoma de MadridCantoblancoMadridSpain School of Environment The University of QueenslandGattonAustralia School of Veterinary Science The University of QueenslandGattonAustralia

Objective:To determine the relationship between teratozoospermia and sperm DNA fragmentation(SDF)in the human ***:This retrospective study included 100 normozoospermic men as a control cohort(abnormal forms>14%),210 patients with a high level of abnormal forms(≤4%)and 65 patients presenting with a moderate level of abnormal forms(>4%to≤14%)based on the World Health Organization *** morphology was assessed using bright field *** DNA fragmentation was assessed using the sperm chromatin dispersion ***-parametric analyses were conducted to examine the relationship between abnormal sperm morphology and sperm DNA fragmentation;receiver operating characteristic(ROC)analyses were conducted to assess sensitivity and specificity of this ***:A correlation analysis revealed that the higher the proportion of abnormal spermatozoa in the ejaculate,the higher the level of SDF(Spearman's Rho=-0.230;P<0.001).Significant differences in the proportion of SDF were found when all cohorts were compared(P<0.001);these significant differences were also retained when the different cohorts were compared *** analysis showed a moderate but significant predictive value for SDF to differentiate patients with different levels of ***:Although analysis of a more continuous range of values for teratozoospermia would help further clarify any causal relationship with SDF,there is clearly a synergistic or coincident affiliation between these variables that needs to be acknowledged by the clinician when interpreting the spermiogram.

关键词： Teratozoospermia Sperm morphology Sperm DNA fragmentation male factor Human reproduction Human fertility

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Can DNA fragmentation of neat or swim-up spermatozoa be used to predict pregnancy following ICSI of fertile oocyte donors？

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Asian Journal of Andrology 2013年第6期15卷 812-818,I0010页

作者： Jaime Gosálvez Pedro Caballero Carmen López-Fernández Leonor Ortega José Andrés Guijarro José Luís FernAndez Stephen D Johnston Rocio Nufiez-Calonge Faculty of Biology Autonomous University of Madrid Cantoblanco Madrid 28049 Spain CIfnica Tambre Madrid 28002 Spain GINIBIC-Complejo Hospitalario Universitario A CoruRa A Corufna15006 Spain School of Agriculture and Food Science The University of Queensland Gatton 4343 Australia

This study compared the potential of assessing sperm DNA fragmentation （SDF） from neat semen and the subsequent swim-up （SU） procedure to predict pregnancy when conducting ICSI of fertile donor oocytes. Infertile females （n=81） were transferred embryos resulting from intracytoplasmic sperm injection （ICSI） of their partner＇s spermatozoa and proven donor oocytes. This model normalized the impact of female factor in putative sperm DNA repair. Semen was blindly assessed for SDF using Halosperm immediately following ejaculation （NS） and after swim-up at the time of ICSI fertilisation. There was a decrease in SDF values of the ejaculated semen sample following the swim-up protocol （P=0.000）. Interestingly, pregnancy could be equally predicted from SDF values derived from either neat or swim-up semen samples. Receiver operator curves and the derived Youden＇s indices determined SDF cutoff values for NS and SU of 24.8% and 17.5%, respectively. Prediction of pregnancy from NS SDF had a sensitivity of 75% and a specificity of 69%, whereas for SU SDF was 78% and 73%, respectively. While increased levels of SDF negatively impact reproductive outcome, we have shown that a reduction in SDF following sperm selection using ICSI with proven donor oocytes is not mandatory for achieving pregnancy. This suggests that a certain level of DNA damage that is not detectable using current technologies could be impacting on the relative success of assisted reproductive technology （ART） procedures. Consequently, we propose a modification of the so called ＇iceberg model＇ as a possible rationale for understanding the role of SDF in reproductive outcome.

关键词： assisted reproductive technology egg donation male factor sperm DNA fragmentation

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Association of polymorphisms in glutathione S-transferase genes （GSTM1, GSTT1, GSTP1） with idiopathic azoospermia or oligospermia in Sichuan, China

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Asian Journal of Andrology 2015年第3期17卷 481-486页

作者： Da-Ke Xiong Key Laboratory of Bio-Resources and Eco-Environment Ministry of Education School of Life Science Institute of Medical Genetics Sichuan University Chengdu China Bio-resource Research and Utilization Joint Key Laboratory of Sichuan and Chongqing Chengdu China Biotechnology Academy of Nanchuan Chongqing China

The reported effects of the glutathione S-transferase （GSTs） genes （GSTM1, GSTTI, and GSTP1） on male factor infertility have been inconsistent and even contradictory. Here, we conducted a case-control study to investigate the association between functionally important polymorphisms in GST genes and idiopathic male infertility. The study group consisted of 361 men with idiopathic azoospermia, 118 men with idiopathic oligospermia, and 234 age-matched healthy fertile male controls. Genomic DNA was extracted from the peripheral blood, and analyzed by polymerase chain reaction and restriction fragment length polymorphism analysis. There was a significant association between the GSTP1 variant genotype （lle/Val ＋ Val/Val） with idiopathic infertility risk （odds ratio [OR]： 1.53; 95% confidence interval [CI]： 1.11-2.11; P = 0.009）. Similarly, a higher risk of infertility was noted in individuals carrying a genotype combination of GSTTI-null and GSTP1 （lle/Val ＋ ValNal）（OR： 2.17; 95% Cl： 1.43-3.31; P = 0.0002）. These results suggest an increased risk of the GSTPI variant genotype （lle/Val ＋ Val/Val） for developing male factor infertility. Our findings also underrate the significance of the effect of GSTM1 and/or GSTT1 （especially the former） in modulating the risk of male infertility in males from Sichuan, southwest China.

关键词： genetic polymorphism glutathione S-transferase idiopathic infertility male factor

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The ability of sperm selection techniques to remove single- or double-strand DNA damage

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Asian Journal of Andrology 2011年第5期13卷 764-768页

作者： Maria Enciso Miriam Iglesias Isabel Galan Jonas Sarasa Antonio Gosalvez Jaime Gosalvez Unidad de Genetica Departamento de Biologia Universidad Aut6noma de Madrid Madrid 28049 Spain Unidad de Reproducci6n Asistida Hospital Quir6n Madrid Madrid 28223 Spain

A wide variety of techniques for the preparation of sperm are currently available, of which the most commonly employed are densitygradient centrifugation （DGC） and swim-up （SUP）. To date, these methods appear to be effective in selecting functional sperm for assisted reproduction techniques （ART）, but they may have negative effects on sperm DNA. In this study, the ability of these semen processing techniques to eliminate spermatozoa containing single- and double-strand DNA damage was assessed by the two-tailed comet assay and the sperm chromatin dispersion test in 157 semen samples from patients seeking assisted reproduction treatment. Our results indicated that SUP and DGC are equally efficient in eliminating spermatozoa containing double-strand DNA damage and sperm with highly damaged （degraded） DNA, as characterized by the presence of both single- and double-strand DNA breaks. However, DGC is more efficient than SUP in selecting spermatozoa that are free from single-strand DNA damage. Future studies should characterise the importance of the various types of DNA damage and examine the sperm processing protocols used in each laboratory to determine their ability to eliminate DNA damage and hence, prevent the potential transmission of genetic mutations via ART.

关键词： density-gradient centrifugation male factor sperm chromatin dispersion sperm DNA fragmentation swim-up

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Future directions of clinical laboratory evaluation ol pregnancy

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Cellular & Molecular Immunology 2014年第6期11卷 582-588页

作者： Kenneth D Beaman Mukesh K Jaiswal Svetlana Dambaeva Alice Gilman-Sachs Department of Microbiology and Immunology Rosalind Franklin University of Medicine and Science North Chicago IL USA

In recent years, our understanding of how the immune system interacts with the developing fetus and placenta has greatly expanded. There are many laboratories that provide tests for diagnosis of pregnancy outcome in women who have recurrent pregnancy loss （RPL） or pre-eclampsia. These tests are based on the premise that immune response to the fetus is equivalent to the adaptive immune response to a transplant. New understanding leads to the concept that the activated innate response is vital for pregnancy and this can result in more effective testing and treatment to prevent an abnormal pregnancy in the future. We describe here only three such areas for future testing： one area involves sperm and semen and factors necessary for successful fertilization; another area would determine conditions for production of growth factors necessary for implantation in the uterus; finally, the last area would be to determine conditions necessary for the vascularization of the placenta and growing fetus by activated natural killer （NK） cells （combinations of killer cell immunoglobulin-like receptor （KIR） family genes with HLA-C haplotypes） that lead to capability of secreting angiogenic growth factors. These areas are novel but understanding their role in pregnancy can lead to insight into how to maintain and treat pregnancies with complicating factors.

关键词： male factor preterm labor recurrent pregnancy loss seminal plasma sperm

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Inter-center variation in the efficiency of sperm DNA damage reduction following density gradient centrifugation

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Natural Science 2013年第7期5卷 15-20页

作者： Carmen López-Fernández Stephen D.Johnston Altea Gosálbez Jose Luís Fernández Juan G.Alvarez Jaime Gosálvez Department of Biology Genetics UnitUniversity Autónoma of MadridMadridSpain School of Agriculture and Food Science The University of QueenslandGattonAustralia Laboratorio de Genética Molecular y Radiobiología Centro Oncológico de GaliciaA CorunaSpain Centro ANDROGEN A CorunaSpain

This was a prospective multicenter study aiming at comparing the efficiency of sperm selection by density gradient centrifugation (DGC) in reducing sperm DNA fragmentation (SDF) in different ART centers. The study was designed using 290 semen samples collected from 10 different ART centers performing artificial insemination, in vitro fertilization and blind assessment of SDF at the University facilities. The results showed that while there was a significant reduction in the SDF levels in sperm isolated from the gradient centrifuged pellet (DGC) compared to neat semen samples (NSS), there was also significant inter-center variability in the efficiency to reduce SDF values by DGC (78.5% to 29.2%). Surprisingly, for some patients, the level of SDF actually increased following sperm selection. The main conclusions derived from this study were that 1) isolation of sperm from the gradient pellet by DGC must be performed using validated, optimized protocols;2) routine comparison of SDF values in NSS semen and in processed sperm after DGC or swim-up must be recommended as part of the internal quality control (QC) of ART laboratories to test the efficacy of sperm processing;and 3) SDF values in processed spermatozoa should be obtained to compare with the pregnancy rate when insemination or fertilization is about to be performed, otherwise, attempts to predict pregnancy outcome from SDF could be biased or are essentially meaningless.

关键词： Assisted Reproductive Technology Fertility Sperm DNA Fragmentation male factor Semen Processing

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