scholarly journals Proteomic Analysis in Seminal Plasma of Fertile Donors and Infertile Patients with Sperm DNA Fragmentation

2020 ◽  
Vol 21 (14) ◽  
pp. 5046
Author(s):  
Alba Fernandez-Encinas ◽  
Agustín García-Peiró ◽  
Javier del Rey ◽  
Jordi Ribas-Maynou ◽  
Carlos Abad ◽  
...  
Keyword(s):  
Male Infertility ◽  
Dna Fragmentation ◽  
Seminal Plasma ◽  
Recurrent Miscarriage ◽  
Semen Analysis ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Plasma Samples ◽  
Sperm Dna ◽  
Dispersion Test

Seminal plasma proteomics studies could represent a new approach for the determination of molecular elements driving male infertility, resulting in a better male infertility characterization. The aim of this study is to investigate proteomic differences in seminal plasma samples from fertile and infertile individuals. For that, semen samples were selected according to semen analysis, clinical pathology, and values of sperm DNA fragmentation (alkaline and neutral Comet assay and Sperm Chromatin Dispersion test). A total of 24 seminal plasma samples classified in four groups were processed: fertile donors (FD), recurrent miscarriage patients (RM), asthenoteratozoospermic patients (ATZ), and asthenoteratozoospermic patients with varicocele (ATZ-VAR). Results obtained by 2D-differential gel electrophoresis (2D-DIGE) revealed 26 spots significantly increased in fertile donors when compared to patient groups. Also, eight spots in the ATZ group and two in the ATZ-VAR group were decreased compared to the other groups. Twenty-eight proteins were identified by mass spectrometry (MS), most of them involved in metabolic and cellular processes and with a catalytic or binding function. Protein–protein interactions through Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) tool suggest that a large part of them were associated with each other. Furthermore, most of them were associated with ubiquitin C, indicating that it could play an important regulation role, resulting in a potential male infertility biomarker.

P–009 A modified sperm chromatin dispersion test, LensHooke® R10, for quick and accurate determination of human sperm DNA fragmentation

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
L S Chang ◽  
H C Lee ◽  
C T Hsu ◽  
H M Tsao ◽  
C C Huang ◽  
...  
Keyword(s):  
Male Infertility ◽  
Dna Fragmentation ◽  
Observer Agreement ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Test Results ◽  
Sperm Dna ◽  
Test Kit ◽  
Dispersion Test ◽  
Sperm Chromatin Dispersion Test

Abstract Study question The performance and efficiency of the LensHooke® R10 test kit were evaluated by the clinical examination for precision, accuracy, and time. Summary answer The LensHooke® R10 based on sperm chromatin dispersion test offers not only quick testing for sperm DNA fragmentation but also reliable and accurate test results. What is known already Sperm chromatin dispersion (SCD) test, one of the most commonly used testing for sperm DNA fragmentation (SDF), can be conducted promptly and without the need for expensive laboratory instruments. However, the main disadvantage of the SCD test is inter-observer variability in categorizing the size of characteristics halos surrounding the core of sperm. Moreover, it takes more than one hour to accomplish whole assay procedures making this testing an inefficient diagnostic tool. These may hinder its broad availability among andrology laboratories or prevent it from being routinely used for the evaluation of male infertility. Study design, size, duration A total of 108 participants was included in this prospective study. Data was collected from the reproductive medicine center between June and December 2020. Participants/materials, setting, methods This study included 108 consecutive male partners of couples attending for assisted reproductive treatment. SDF was simultaneously tested by using LensHooke® R10 (R10) and Halosperm® G2 (G2) respectively. We evaluated the correlation and agreement between two SCD-based test kits. The repeatability and reproducibility of the SCD kits were assessed by intra-and inter-observer agreement experiments. The sensitivity, specificity, positive predictive value, negative predictive value for the R10 was determined by receiver operator characteristics (ROC) curve analysis. Main results and the role of chance The R10 produced more clear sperm core and dispersed chromatin, therefore highly recognizable images can be easily and accurately categorized when scoring of SDF. It took 50% less time for SDF testing by the R10 compared to the G2 (38.26 ± 9.85 minutes vs. 76.52 ± 19.7 minutes, P < 0.0001). The SDF% results showed a strong correlation for the R10 and G2 with Spearman’s coefficients of rank correlation (rho) above 0.8 (P < 0.0001, N = 108). The R10 showed 89.8% accuracy with 87.9% sensitivity, 90.8% specificity, 82.9% PPV, and 93.7% NPV on the measurement of SDF% at the threshold value of 22%. Intraclass correlation coefficients (ICC) >0.9 showed a strong agreement between two observers on the testing of SDF using the R10. ICC >0.9 showed a high intra-observer agreement within 4 repeated testing on SDF using the R10. The R10 showed an intra-observer’s precision of coefficient variation, CV < 10% for SDF%. In addition, SDF% test results obtained by the R10 for asthenospermic (31.8% ± 16.7%), teratospermic (22.9% ± 14.4%), and oligoasthenoteratozoospermic samples (36.6% ± 14.4%) were significantly higher than that observed in normozoospermic samples (15.3% ± 10.2%, p < 0.05), was comparable with the G2. Limitations, reasons for caution The sample size of 4 semen specimens used to evaluate the intra-and inter-observer agreement was a limitation. Besides, evaluating the relationship between the SDF and clinical outcome of ART is necessary for further study. Wider implications of the findings: The new in vitro diagnostics reagent, LensHooke® R10, is a simple and quick test kit that offers reliable and accurate test results of sperm DNA fragmentation, can be routinely used in male infertility evaluation. Trial registration number CS2–20012

The assessment of sperm DNA fragmentation in the saltwater crocodile (Crocodylus porosus)

2017 ◽  
Vol 29 (3) ◽  
pp. 630 ◽  
Author(s):  
S. D. Johnston ◽  
C. López-Fernández ◽  
F. Arroyo ◽  
J. L. Fernández ◽  
J. Gosálvez
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Fragmented Dna ◽  
Sperm Dna ◽  
Saltwater Crocodile ◽  
Crocodylus Porosus ◽  
Dispersion Test ◽  
Sperm Nuclei

Herein we report a method of assessing DNA fragmentation in the saltwater crocodile using the sperm chromatin dispersion test (SCDt) after including frozen–thawed spermatozoa in a microgel (Halomax; Halotech DNA, Madrid, Spain). Following controlled protein depletion, which included a reducing agent, sperm nuclei with fragmented DNA showed a homogeneous and larger halo of chromatin dispersion with a corresponding reduced nucleoid core compared with sperm with non-fragmented DNA. The presence of DNA damage was confirmed directly by incorporation of modified nucleotides using in situ nick translation (ISNT) and indirectly by studying the correlation of the SCDt with the results of DNA damage visualisation using a two-tailed comet assay (r = 0.90; P = 0.037). Results of the SCDt immediately following thawing and after 5 h incubation at 37°C in order to induce a range of DNA damage revealed individual crocodile differences in both the baseline level of DNA damage and DNA longevity.

P–068 CatSper4 cation channel expression is low in male infertility and is affected by cryopreservation

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
N Kilic ◽  
T İrez ◽  
N Dayiolu
Keyword(s):  
Male Infertility ◽  
Dna Fragmentation ◽  
Semen Analysis ◽  
Middle Part ◽  
Cation Channel ◽  
The Other ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Negative Effects ◽  
Sperm Dna

Abstract Study question Is CatSper4 expression in sperm related to functional parameters and does cryopreservation affect CatSper4 expression? Summary answer In this study, it was aimed to investigate whether CatSper4 has a relationship with sperm parameters and is CatSper 4 affected by cryopreservation. What is known already CatSper membrane channels, known as cation channels, are thought to play an important role in the insufficiency of sperm physiology, acrosome reaction, and chemotaxis movement. There is no study on cation channel distribution in an infertile male patient. In addition, studies conducted in recent years have shown that cryopreservation techniques have negative effects on sperm DNA, but there is no analysis in the literature regarding the effects of cryopreservation on CatSper4 ion channel proteins. Study design, size, duration Samples of the patients who applied to the Andrology laboratory in the Medical Park Hospital IVF unit between March 1 and June 1 in 2020 were included in the study. Also, patients with no family history of no genetic anomalies , no varicocele and azoospermia were included.The study were divided into 4 groups in accordance with the male infertility guideline of the European Association of Urology as normozoospermic (control group), the asthenoteratozoospermia, teratozoospermia, and oligoastenotheratozoospermia. Participants/materials, setting, methods In this prospective study, semen analysis, DNA fragmentation, and CatSper 4 by IHC of control group patients with normospermia (n = 40) and oligospermia(n = 50), asthenospermia(n = 40), and teratozoospermia(n = 38) patients were compared and differences resulting from cryopreservation were evaluated by Wilcoxon signed Ranks Test. Main results and the role of chance It was observed that CatSper4 protein positivity was localized in the middle part of the sperm and it was statistically higher in the normozoospermic patient group compared to the other groups (p = 0,01). When the positivity values of CatSper4 protein before and after freezing were compared in the groups, it was seen that the values decreased (p = 0,001,p=0,01). Sperm DNA fragmentation was found to be lowest in normospermia and statistically significantly higher in other groups. Cryopreservation application increased DNA fragmentation in all groups (p < 0,001 , p < 0,01). Limitations, reasons for caution Unfortunately, embryo screening in patients with low CatSper4 expression is not available in the present study. Soon we plan to screen a broader clinical pregnancy series and present the IVF results associated with CatSper4. Wider implications of the findings: Our study indicated that, CatSper4 expression is quite high in normospermia when compared with the other groups, particularly oligoasthenoteratozoospermia and asthenoteratozoospermia. There are almost no studies on this subject in the literature, and we think that it should be studied in larger patient groups and in unexplained infertile cases. Trial registration number Not applicable

Characterization of Nuclease Activity in Human Seminal Plasma and its Relationship to Semen Parameters, Sperm DNA Fragmentation and Male Infertility

2016 ◽  
Vol 195 (1) ◽  
pp. 213-219 ◽  
Author(s):  
Alba Fernandez-Encinas ◽  
Agustí García-Peiró ◽  
Jordi Ribas-Maynou ◽  
Carlos Abad ◽  
María José Amengual ◽  
...  
Keyword(s):  
Male Infertility ◽  
Dna Fragmentation ◽  
Seminal Plasma ◽  
Nuclease Activity ◽  
Semen Parameters ◽  
Sperm Dna Fragmentation ◽  
Human Seminal Plasma ◽  
Sperm Dna

Sperm DNA fragmentation: causes and identification

Zygote ◽  
2019 ◽  
Vol 28 (1) ◽  
pp. 1-8 ◽  
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.

Sperm chromatin dispersion test (SCDt) for the assessment of sperm DNA fragmentation in black tiger prawn, Penaeus monodon

Aquaculture ◽  
2018 ◽  
Vol 491 ◽  
pp. 281-288 ◽  
Author(s):  
Tianyi Feng ◽  
Jamie Gosálvez ◽  
Carmen Lopez-Fernandez ◽  
Francisca Arroyo ◽  
Brian Paterson ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Penaeus Monodon ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna ◽  
Black Tiger Prawn ◽  
Tiger Prawn ◽  
Dispersion Test ◽  
Sperm Chromatin Dispersion Test

Assessment of Sperm DNA Fragmentation in Stallion (Equus caballus) and Donkey (Equus asinus) Using the Sperm Chromatin Dispersion Test

2009 ◽  
Vol 44 (5) ◽  
pp. 823-828 ◽  
Author(s):  
EI Cortés-Gutiérrez ◽  
F Crespo ◽  
C Serres-Dalmau ◽  
AL Gutiérrez de las Rozas ◽  
MI Dávila-Rodríguez ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Equus Caballus ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Equus Asinus ◽  
Sperm Dna ◽  
Dispersion Test ◽  
Sperm Chromatin Dispersion Test
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