Development of Normal Reference Values for Seminal Reactive Oxygen Species and Their Correlation With Leukocytes and Semen Parameters in a Fertile Population

BACKGROUND Infertility is defined as the inability to conceive after at 1 year of regular unprotected intercourse. Male contributes to almost half of infertility cases and in almost 30 % of cases, no definite aetiology is identified, and hence, male infertility is labelled idiopathic in these cases. Oxidative energy production mechanisms are almost always accompanied by reactive oxygen species (ROS), generation whose too much concentrations can lead to extensive protein damage and cytoskeletal modifications and inhibit cellular mechanisms. A number of laboratory techniques have been developed to evaluate oxidative stress by measuring ROS level in the semen. In recent times antioxidant supplements have been proposed as useful agents to increase the scavenging capacity of seminal plasma, controversy still surrounds their actual clinical utility. METHODS 34 male patients were included in this study. Reactive oxygen species detection was done by Flowcytometry using dichloroflurosecindiacetate (DCFH-DA). RESULTS The ROS in the patient group was found to be significantly higher 29.821 (5.6300 than the control group 22.162 (1.6331 having p value < 0.001). The ROS (29.821 ± 5.6300) was found to be significantly reduced after 3 months of antioxidant therapy which got reduced to 19.893 ± 4.2299 respectively. CONCLUSIONS Our study demonstrates that infertile men have significantly higher level of ROS (as measured by flowcytometry) & lower sperm count (oligospermia), decreased progressive & total motility and increased immotile sperms as compared to healthy fertile men. This study further proves that antioxidant therapy based on a combination of carnitine, zinc, coq10, lycopene and vitamin C & E for 3 months is associated with a decrease of ROS as measured by flowcytometry & a variable degree of improvement in above mentioned semen parameters. KEYWORDS Reactive Oxygen Species, Male Infertility

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Low amounts of mitochondrial reactive oxygen species define human sperm quality

Reproduction ◽

10.1530/rep-13-0644 ◽

2014 ◽

Vol 147 (6) ◽

pp. 817-824 ◽

Cited By ~ 10

Author(s):

Mónica Marques ◽

Ana Paula Sousa ◽

Artur Paiva ◽

Teresa Almeida-Santos ◽

João Ramalho-Santos

Keyword(s):

Reactive Oxygen Species ◽

Assisted Reproduction ◽

Density Gradient Centrifugation ◽

Sperm Quality ◽

Gradient Centrifugation ◽

Reactive Oxygen ◽

Human Sperm ◽

Semen Parameters ◽

Oxygen Species ◽

Male Gametes

We have applied the mitochondria-specific superoxide fluorescent probe MitoSOX Red (MitoSOX) to detect mitochondria-specific reactive oxygen species (mROS) production in human sperm samples using flow cytometry. We show that human ejaculates are heterogeneous in terms of mROS production, with three subpopulations clearly detectable, comprising sperm that produce increasing amounts of mROS (MitoSOX−, MitoSOX+, and MitoSOX++). The sperm subpopulation producing the lowest amount of mROS represented the most functional subset of male gametes within the ejaculate, as it was correlated with the highest amount of live and non-apoptotic sperm and increased both in samples with better semen parameters and in samples processed by both density-gradient centrifugation and swim-up, both known to select for higher quality sperm. Importantly, the MitoSOX− subpopulation was clearly more prevalent in samples that gave rise to pregnancies following assisted reproduction. Our work, therefore, not only describe discreet human sperm heterogeneity at the mROS level but also suggests that mROS may represent a strategy to both evaluate sperm samples and isolate the most functional gametes for assisted reproduction.Free Portuguese abstractA Portuguese translation of this abstract is freely available athttp://www.reproduction-online.org/content/147/6/817/suppl/DC1

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Male infertility and the effect of seminal and urinary infections on sperm DNA fragmentation, reactive oxygen species and WHO semen parameters

European Urology ◽

10.1016/s0302-2838(21)00886-1 ◽

2021 ◽

Vol 79 ◽

pp. S694

Author(s):

C.L.T. Ho ◽

D.R. Vaughan-Constable ◽

J. Ramsay ◽

C. Jayasena ◽

T. Tharakan ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Male Infertility ◽

Dna Fragmentation ◽

Reactive Oxygen ◽

Semen Parameters ◽

Sperm Dna Fragmentation ◽

Oxygen Species ◽

Urinary Infections ◽

Sperm Dna

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DEFINING REFERENCE VALUES FOR SEMINAL REACTIVE OXYGEN SPECIES (ROS) IN A POPULATION OF INFERTILE MEN USING RECEIVER OPERATING CHARACTERISTIC (ROC) CURVE

The Journal of Urology ◽

10.1016/s0022-5347(08)61752-8 ◽

2008 ◽

Vol 179 (4S) ◽

pp. 597-598

Author(s):

Rakesh Sharma ◽

Reda Mahfouz ◽

Nisarg Desai ◽

Kelly S Athayde ◽

Edmund Sabanegh ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Receiver Operating Characteristic ◽

Reference Values ◽

Roc Curve ◽

Operating Characteristic ◽

Reactive Oxygen ◽

Oxygen Species ◽

Infertile Men ◽

Receiver Operating

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A Comparison Between Two Assays for Measuring Seminal Oxidative Stress and their Relationship with Sperm DNA Fragmentation and Semen Parameters

Genes ◽

10.3390/genes10030236 ◽

2019 ◽

Vol 10 (3) ◽

pp. 236 ◽

Cited By ~ 18

Author(s):

Sheryl Homa ◽

Anna Vassiliou ◽

Jesse Stone ◽

Aideen Killeen ◽

Andrew Dawkins ◽

...

Keyword(s):

Oxidative Stress ◽

Reactive Oxygen Species ◽

Dna Fragmentation ◽

Recurrent Miscarriage ◽

Reactive Oxygen ◽

Semen Parameters ◽

Sperm Chromatin ◽

Sperm Dna Fragmentation ◽

Oxygen Species ◽

Sperm Dna

Oxidative stress (OS) is a significant cause of DNA fragmentation and is associated with poor embryo development and recurrent miscarriage. The aim of this study was to compare two different methods for assessing seminal OS and their ability to predict sperm DNA fragmentation and abnormal semen parameters. Semen samples were collected from 520 men attending for routine diagnostic testing following informed consent. Oxidative stress was assessed using either a chemiluminescence assay to measure reactive oxygen species (ROS) or an electrochemical assay to measure oxidation reduction potential (sORP). Sperm DNA fragmentation (DFI) and sperm with immature chromatin (HDS) were assessed using sperm chromatin structure assay (SCSA). Semen analysis was performed according to WHO 2010 guidelines. Reactive oxygen species sORP and DFI are negatively correlated with sperm motility (p = 0.0012, 0.0002, <0.0001 respectively) and vitality (p < 0.0001, 0.019, <0.0001 respectively). The correlation was stronger for sORP than ROS. Reactive oxygen species (p < 0.0001), sORP (p < 0.0001), DFI (p < 0.0089) and HDS (p < 0.0001) were significantly elevated in samples with abnormal semen parameters, compared to those with normal parameters. Samples with polymorphonuclear leukocytes (PMN) have excessive ROS levels compared to those without (p < 0.0001), but sORP and DFI in this group are not significantly increased. DNA fragmentation was significantly elevated in samples with OS measured by ROS (p = 0.0052) or sORP (p = 0.004). The results demonstrate the multi-dimensional nature of oxidative stress and that neither assay can be used alone in the diagnosis of OS, especially in cases of leukocytospermia.

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