Cementing the relationship between conventional and advanced semen parameters

Abstract Background Affordable conventional semen analysis remains a fundamental procedure to be performed routinely during the diagnosis of male infertility. Advanced semen analyses provide valuable clinical insights in treatment-related decision-making, but these are highly expensive and lack universal standardization. This study aimed at determining the relationship between conventional semen parameters, measured with assistance of computer-aided sperm analysis (CASA), and a set of advanced semen tests. Basic semen analysis (n = 124) was performed according to the World Health Organization (WHO) guidelines. Sperm DNA fragmentation and intracellular superoxide (O2−•) levels were assessed by flow cytometry. Seminal plasma thiobarbituric acid reactive substances (TBARS) levels as well as superoxide dismutase (SOD) and catalase (CAT) activity were measured by spectrophotometry. Spearman’s rank correlation coefficient was used, with significance set at p < 0.05. Results Semen pH correlated negatively with TBARS (p < 0.01). The proportions of total and progressively motile as well as rapid spermatozoa correlated positively with CAT activity (p < 0.05). Sperm viability correlated negatively with both O2−• (p < 0.05) and DNA fragmentation (p = 0.01), while normal morphology correlated negatively with O2−• levels (p < 0.05) and positively with CAT activity (p < 0.05). Straight-line velocity (VCL) and average-path velocity (VAP) correlated negatively with both O2−• (p < 0.01) and TBARS (p < 0.01). Amplitude of lateral head displacement (ALH) correlated negatively with O2−• (p < 0.01) and DNA fragmentation (p < 0.01), while its correlation with SOD activity was positive (p < 0.05). Conclusion The results obtained from this study support the validity of some CASA parameters as sensitive indicators of changes in sperm oxidative status and DNA integrity. Predicting advanced from conventional parameters through the building of linear regression models should be considered for future studies.

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Relationships between sperm DNA integrity and bulk semen parameters in Bulgarian patients with varicocele

Archivio Italiano di Urologia e Andrologia ◽

10.4081/aiua.2019.2.125 ◽

2019 ◽

Vol 91 (2) ◽

Author(s):

Viktor Alargkof ◽

Larissa Kersten ◽

Romil Stanislavov ◽

Zdravko Kamenov ◽

Panagiotis Nikolinakos

Keyword(s):

Dna Fragmentation ◽

Semen Analysis ◽

Human Reproduction ◽

Sperm Parameters ◽

Semen Parameters ◽

Control Group ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Abnormal Sperm ◽

Sperm Dna

Objective: This exploratory retrospective study aimed to compare the level of Sperm DNA Fragmentation (SDF) and investigate its association with bulk semen parameters, for the first time in Bulgarian patients with varicocele, using a distinct methodology. Material and methods: Standard semen analysis was performed according to the 2010 criteria of the European Society of Human Reproduction and Embryology - Nordic Association for Andrology (ESHRE-NAFA-2010) and DNA fragmentation was assessed using the Halosperm® kit. The total sample included 28 males: the control group consisted of men with normal genital examination and unknown fertility (n = 10), group one consisted of men with varicocele, normozoospermia and DNA fragmentation > 15% (n = 9) and group two consisted of men with varicocele, abnormal sperm parameters and DNA fragmentation > 15% (n = 9). Results: DNA fragmentation was found to be higher in patients with abnormal sperm parameters (43.78 ± 30.78) compared to the normozoospermic group (21.22 ± 3.93) (p = 0.008). In normozoospermic patients, no statistically significant correlations were observed between SDF and bulk semen parameters. In patients with abnormal sperm parameters, DNA fragmentation exhibited significant very strong negative association with motility (a+b), vitality and typical morphology (p < 0.001). Conclusions: DNA integrity assays could be used for a better evaluation and management of male infertility, particularly in normozoospermic varicocele patients.

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Correlation among isolated teratozoospermia, sperm DNA fragmentation and markers of systemic inflammation in primary infertile men

PLoS ONE ◽

10.1371/journal.pone.0251608 ◽

2021 ◽

Vol 16 (6) ◽

pp. e0251608

Author(s):

Luigi Candela ◽

Luca Boeri ◽

Paolo Capogrosso ◽

Walter Cazzaniga ◽

Edoardo Pozzi ◽

...

Keyword(s):

Linear Regression ◽

Dna Fragmentation ◽

Semen Analysis ◽

Linear Regression Analysis ◽

Semen Parameters ◽

Sperm Dna Fragmentation ◽

Linear Regression Models ◽

Inflammatory Parameters ◽

Infertile Men ◽

Sperm Dna

Aim To assess the prevalence of isolated teratozoospermia (iTZS) in a cohort of infertile and fertile men; explore the relationship between iTZS, inflammatory parameters and sperm DNA fragmentation index (SDF) in the same cohort. Materials and methods 1824 infertile men and 103 fertile controls. Semen analysis, the neutrophil-to-lymphocyte ratio (NLR) and serum hormones were investigated. DFI was tested in infertile men only. According to 2010 WHO semen analysis, patients were categorized in 3 sub-groups of isolated sperm defects: isolated oligozoospermia (iOZS), isolated asthenozoospermia (iAZS) and iTZS. Descriptive statistics and linear regression models tested the association between clinical variables and inflammatory markers. Results Among infertile men, iAZS, iTZS, and iOZS were found in 13.9%, 11.9% and 4.1% participants, respectively. iTZS was found in 37 (35.9%) fertile men. Infertile men with iTZS had higher NLR values than those with iOZS, iAZS and men with normal semen parameters (all p<0.001). FSH and LH were higher and inhibin B lower in iOZS infertile men compared to all other groups (p≤0.001). Hormonal characteristics were similar between iTZS infertile and fertile men. Similarly, iTZS infertile men had higher SDF than all other groups (all p<0.001). Infertile men with iTZS had higher NLR values than fertile men with iTZS (p<0.01). Linear regression analysis showed that, in infertile men, iTZS was associated with SDF and NLR (all p≤0.01). Conclusions iTZS was found in 11.9% of infertile men but it was even more prevalent in fertile controls. Infertile men with iTZS had higher NLR than fertile controls and increased SDF values than infertile participant with iAZS, iOZS, or normal semen parameters. No differences in hormonal characteristics were found between infertile and fertile men with iTZS.

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Sperm DNA fragmentation: causes and identification

Zygote ◽

10.1017/s0967199419000595 ◽

2019 ◽

Vol 28 (1) ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Thais Rose dos Santos Hamilton ◽

Mayra Elena Ortiz D’Ávila Assumpção

Keyword(s):

Dna Fragmentation ◽

Semen Analysis ◽

Sperm Chromatin ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Functional Capability ◽

Sperm Dna Integrity ◽

Sperm Dna ◽

Fertility Disorders ◽

Chromatin Integrity

SummarySperm DNA fragmentation is referred to as one of the main causes of male infertility. Failures in the protamination process, apoptosis and action of reactive oxygen species (ROS) are considered the most important causes of DNA fragmentation. Action of ROS or changes in sperm protamination would increase the susceptibility of sperm DNA to fragmentation. Routine semen analysis is unable to estimate sperm chromatin damage. Sperm DNA integrity influences sperm functional capability, therefore tests that measure sperm DNA fragmentation are important to assess fertility disorders. Actually, there is a considerable number of methods for assessing sperm DNA fragmentation and chromatin integrity, sperm chromatin stability assay (SCSA modified), sperm chromatin dispersion (SCD), comet assay, transferase dUTP nick end labelling (TUNEL); and protamine evaluation in sperm chromatin assay, such as toluidine blue, CMA3, protamine expression and evaluation of cysteine radicals. This review aims to describe the main causes of sperm DNA fragmentation and the tests commonly used to evaluate sperm DNA fragmentation.

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Low-dose Methotrexate Therapy Does Not Affect Semen Parameters and Sperm DNA

Inflammatory Bowel Diseases ◽

10.1093/ibd/izab205 ◽

2021 ◽

Author(s):

Anne Grosen ◽

Emanuelle Bellaguarda ◽

Jacob Nersting ◽

Christian Lodberg Hvas ◽

Ingela Liljeqvist-Soltic ◽

...

Keyword(s):

Healthy Volunteers ◽

Dna Fragmentation ◽

Low Dose ◽

Inflammatory Diseases ◽

Methotrexate Therapy ◽

Semen Parameters ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Fragmentation Index ◽

Sperm Dna

Abstract Background Methotrexate is widely used in inflammatory diseases during the patients’ reproductive years. The effect on male fertility and sperm DNA integrity is largely unknown. We evaluated sperm DNA integrity and basic semen parameters according to the World Health Organization (WHO) in male patients with inflammatory diseases treated with methotrexate. Methods Semen samples from 14 patients on low-dose maintenance methotrexate were compared with samples from 40 healthy volunteers. Further, 5 patients delivered samples on and off methotrexate therapy for paired comparison. Sperm DNA fragmentation index (DFI), concentration, motility, and morphology were evaluated. Blood sex hormones and methotrexate levels were measured in blood and semen. Results DNA fragmentation index in methotrexate-treated patients was comparable with that in healthy volunteers (DFI, 11.5 vs 15.0; P = .06), and DFI did not change significantly on and off methotrexate in the paired samples (DFI, 12.0 vs 14.0; P = 0.35). Sperm concentration, motility, and morphology did not differ between men treated with methotrexate and healthy volunteers. Sperm progressive motility increased off therapy compared with on therapy (65.0% vs 45.0%, P = .04), but all fluctuations in progressive motility were within the WHO reference interval. All methotrexate polyglutamates1-5 were detected in blood, but only methotrexate polyglutamate1 in semen. Serum testosterone was unaffected by methotrexate therapy. Conclusions Patients treated with low-dose methotrexate have a sperm quality comparable with that of healthy volunteers, and methotrexate treatment does not increase sperm DNA fragmentation. This study does not support cryopreservation of semen before treatment initiation nor a 3-month methotrexate-free interval prior to conception.

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Acridine Orange and Flow Cytometry: Which Is Better to Measure the Effect of Varicocele on Sperm DNA Integrity?

Advances in Urology ◽

10.1155/2015/814150 ◽

2015 ◽

Vol 2015 ◽

pp. 1-6 ◽

Cited By ~ 9

Author(s):

Essam-Elden M. Mohammed ◽

Eman Mosad ◽

Asmaa M. Zahran ◽

Diaa A. Hameed ◽

Emad A. Taha ◽

...

Keyword(s):

Flow Cytometry ◽

Acridine Orange ◽

Pregnancy Outcome ◽

Dna Fragmentation ◽

Chromatin Condensation ◽

Semen Parameters ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Sperm Dna Damage ◽

Sperm Dna

We evaluated the effect of varicocelectomy on semen parameters and levels of sperm DNA damage in infertile men. A total of 75 infertile men with varicocele and 40 fertile men (controls) were included in this study. Semen analysis and sperm DNA damage expressed as the DNA fragmentation index using acridine orange staining and chromatin condensation test by flow cytometry were assessed before and 6 months after varicocelectomy. The patients were also followed up for 1 year for pregnancy outcome. Semen parameters were significantly lower in varicocele patients compared to controls (P<0.05). Mean percentages of sperm DNA fragmentation and sperm DNA chromatin condensation in patients were significantly higher than those in controls (P<0.05). After varicocelectomy, sperm DNA fragmentation improved significantly, whereas sperm chromatin condensation was not significantly changed. In 15 out of 75 varicocele patients, clinical pregnancy was diagnosed; those with positive pregnancy outcome had significant improvement in sperm count, progressive sperm motility, and sperm DNA fragmentation, but there was no significant difference in sperm DNA condensation compared to negative pregnancy outcome patients. We concluded from this study that acridine orange stain is more reliable method than flow cytometry in the evaluation of sperm DNA integrity after varicocelectomy.

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Is there a connection with basic sperm parameters and age?

Experimental and Сlinical Urology ◽

10.29188/2222-8543-2020-13-4-58-64 ◽

2020 ◽

Vol 13 (4) ◽

pp. 58-64

Author(s):

A.I. Ryzhkov ◽

◽

I.S. Shormanov ◽

S.Yu. Sokolova ◽

◽

...

Keyword(s):

Dna Fragmentation ◽

Male Fertility ◽

Reproductive Technologies ◽

Semen Parameters ◽

Sperm Dna Fragmentation ◽

Who Guidelines ◽

Negative Relation ◽

Sperm Analysis ◽

Mobile Forms ◽

Sperm Dna

Introduction. TStandard sperm examination, performed according to WHO guidelines, remains the main method of assessment of male fertility. At the same time, this research method has a number of significant limitations, including a low predictive value regarding the outcomes of assisted reproductive technologies (ART) programs and the outcome of naturally occurring pregnancies. The limitations of standard sperm examination dictate the need for additional methods for assessing male fertility. The most promising and widely used test is the assessment of the level of sperm DNA fragmentation. Aim. To evaluate the associations between the levels of sperm DNA fragmentation and the age of the patients along with the following parameters used as a part of standard sperm analysis: semen volume, total number of spermatozoa, percentages of progressively-mobile, non-progressively-mobile and immobile forms, percentages of morphologically normal forms and the forms bearing head defects within the structure of total number of morphological anomalies, as well as semen leukocyte count. Materials and methods. Study materials were the examination results from 121 males aged from 21 to 53 years old (mean age 32.7±4.5 years old), undergoing an examination within the Clinical Institution «Mother and Child – Yaroslavl» during a time period from January 2019 until April 2020. Standard sperm analysis procedures were carried out according to latest edition of WHO Guidelines (2010). The determinations of germ cell DNA fragmentation levels were performed using the TUNEL method. Results. The test results have revealed a weak negative relation between the sperm DNA fragmentation (%) and the percentage of progressively-mobile forms (%) – r = -0.26 (p <0.01). The correlation of sperm DNA fragmentation with age and other parameters was considered statistically insignificant. The second stage of the analysis have demonstrated that the increased degree sperm DNA fragmentation is 1.8-fold more often found in the patients having signs of astenozoospermia (23.6%) comparing to the patients showing normal degree of spermatozoa mobility (13.1%), (p <0.05). Conclusions. The level of sperm DNA fragmentation correlates with the percentage of progressively-mobile forms of spermatozoa (negative relation) and no correlations were found with other semen parameters and with the age. The rates of increased levels of sperm DNA fragmentation are 1.8-fold more often found in astenozoospermia patients comparing to the patients showing normal degrees of spermatozoa mobility.

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The relationship between genitourinary microorganisms and oxidative stress, sperm DNA fragmentation and semen parameters in infertile men

Andrologia ◽

10.1111/and.14322 ◽

2021 ◽

Author(s):

Carmen Lok Tung Ho ◽

Daniella R. Vaughan‐Constable ◽

Jonathan Ramsay ◽

Channa Jayasena ◽

Tharu Tharakan ◽

...

Keyword(s):

Oxidative Stress ◽

Dna Fragmentation ◽

Semen Parameters ◽

Sperm Dna Fragmentation ◽

Infertile Men ◽

Sperm Dna ◽

The Relationship ◽

And Oxidative Stress

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Are semen quality parameters sufficient for biomonitoring spermatozoa DNA integrity and oxidatively damaged DNA

Biomonitoring ◽

10.1515/bimo-2015-0004 ◽

2015 ◽

Vol 2 (1) ◽

Author(s):

Hueiwang Anna Jeng ◽

Ruei-Nian Li ◽

Wen-Yi Lin

Keyword(s):

Dna Damage ◽

Dna Fragmentation ◽

Semen Quality ◽

Quality Parameters ◽

Semen Parameters ◽

Sperm Chromatin ◽

Dna Integrity ◽

Phase System ◽

Sperm Dna Fragmentation ◽

Sperm Dna

Abstract:The present study aimed to investigate the relationship between semen quality parameters and DNA integrity, and determine whether semen quality parameters could serve as a reliable biomarker for monitoring sperm DNA damage. Conventional semen parameters from a total of 202 male human subjects were analyzed. DNA fragmentation and 8-oxo-7,8-dihydro-2′- deoxyguanosine (8-oxoGuo) were used to assess sperm DNA integrity. DNA fragmentation was analyzed by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and sperm chromatin structure assay (SCSA), while 8-oxodGuo was quantified by the liquid chromatography/tandem mass spectrometry (LC-MS/MS) coupled with an on-line solid phase system. The levels of 8-oxodGuo levels in sperm were related to the percentages of DNA fragmentation measured by both the TUNEL and SCSA (r = 0.22, p = 0.048; r = 0.12, p = 0.039). Sperm vitality, motility and morphology from all of the participants exhibited a weak correlation with the levels of 8-oxodGuo and the percentages of DNA fragmentation. Semen quality parameters may be independent of the formation of DNA fragmentation and oxidative adducts in sperm. Semen quality parameters may be insufficient to monitor sperm DNA fragmentation and oxidative damage. DNA damage in sperm is recommended to be included in routine measurements.

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Sperm DNA fragmentation: etiology, pathogenesis, the influence on reproductive function

Urologicheskie vedomosti ◽

10.17816/uroved44804 ◽

2020 ◽

Vol 10 (4) ◽

pp. 337-345

Author(s):

Maxim N. Korshunov ◽

Ekaterina S. Korshunova ◽

Sergey P. Darenkov

Keyword(s):

Dna Fragmentation ◽

Reproductive Function ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Dna Breaks ◽

Sperm Dna Damage ◽

Sperm Dna ◽

Recurrent Pregnancy Losses ◽

Clinical Interest ◽

The Relationship

The literature review of sperm DNA fragmentation is evaluated. Russian and foreign literary data over the past 10 years, including fundamental researching of the evaluation of the gametes genome integrity, are analyzed. The main etiological factors and the possible reasons of the DNA breaks formation on the different stages of spermatogenesis are described. The influence of the sperm oxidative stress reaction to the DNA integrity is analyzed. The relationship between DNA fragmentation of spermatozoa pregnancy and live birth rates in the assisted reproductive techniqueare noted. Risk of the recurrent pregnancy losses in male infertility cases with the sperm DNA damage is presented. Review confirms the significant prognostic value of sperm DNA fragmentation detection in infertility cases. Further studies in evaluation of pathogenesis of sperm DNA have a clinical interest to reproductive health physicians.

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