Sperm DNA damage is related to field fertility of semen from young Norwegian Red bulls

2006 ◽  
Vol 18 (7) ◽  
pp. 781 ◽  
Author(s):  
K. E. Waterhouse ◽  
T. Haugan ◽  
E. Kommisrud ◽  
A. Tverdal ◽  
G. Flatberg ◽  
...  
Keyword(s):  
Dna Damage ◽  
Plasma Membrane ◽  
Sperm Quality ◽  
Sperm Chromatin ◽  
Sperm Dna Damage ◽  
Terminal Deoxynucleotide Transferase ◽  
Fertility Data ◽  
Acrosome Integrity ◽  
Group 2 ◽  
Group 1

Flow cytometry was utilised for the first time to independently measure five sperm parameters of individual spermatozoa of bull ejaculates to differentiate between outcome successes after artificial insemination (AI). These parameters included plasma membrane and acrosome integrity, mitochondrial functionality and DNA damage measured by sperm chromatin structure assay (SCSA) and terminal deoxynucleotide transferase-mediated dUTP nick end labelling (TUNEL) assays. For each parameter, results of 142 ejaculates (30 bulls) were ranked into three groups according to their flow cytometric measures: (1) ejaculates with the 25% lowest measures; (2) the 50% middle measures; and (3) the 25% highest measures. In total, 20 272 first-service inseminations (18 × 106 spermatozoa per AI dose) were performed, where fertility was defined as non-return within 60 days after first insemination. While plasma membrane and acrosome integrity, and mitochondrial functionality were not significantly related to fertility, data from SCSA and TUNEL assays were significantly associated with fertility. Ejaculates in SCSA group 1 had higher odds of AI success (1.07, 95% CI = 1.02–1.12), whereas those in group 3 had lower odds of AI success (0.94, 95% CI = 0.89–0.99), compared with the average odds of all three groups. Ejaculates in group 2 did not have significantly higher odds of AI success compared with the average odds. For TUNEL-positive spermatozoa, the odds of AI success was higher in group 1 compared with the average odds (1.10, 95% CI = 1.02–1.13), whereas odds of AI success in groups 2 and 3 were not significant compared with the average odds. In conclusion, despite the high number of spermatozoa per AI dose from high-quality bulls, both SCSA and TUNEL assays were valuable measures in this study for evaluating sperm quality in relation to fertility after AI.

scholarly journals Ejaculate Oxidative Stress Is Related with Sperm DNA Fragmentation and Round Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Valeria Maria Iommiello ◽  
Elena Albani ◽  
Alessandra Di Rosa ◽  
Alessandra Marras ◽  
Francesca Menduni ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Dna Fragmentation ◽  
Sperm Quality ◽  
Reproductive Capacity ◽  
World Health ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna Damage ◽  
Sperm Dna

Oxidative stress (OS) plays an essential role in male infertility aetiology by affecting sperm quality, function, and also the integrity of sperm DNA. The assessment of oxidative stress in semen may be an important tool to improve the evaluation of sperm reproductive capacity. The purpose of this study was the evaluation of any possible relation between the unbalance of oxidative stress caused by superoxide anion in the ejaculate with the presence of sperm DNA fragmentation and high concentration of round cells. 56 semen samples from males from couples suffering from infertility were evaluated according to World Health Organisation (WHO) 2010 guidelines. Oxidative stress levels from N1 (low) to N4 (high) were assessed in ejaculates using oxiSperm; DFI (sperm DNA fragmentation index) as assessed by the SCSA (Sperm Chromatin Structure Assay) was used for evaluation of sperm chromatin integrity. Our data show that high oxidative stress (N3-N4 levels) correlated positively with aDFI≥30%(P=0.0379)and round cells≥1.500.000/mL(P=0.0084). In conclusion, OS increases sperm DNA damage. Thus evaluation of semen OS extent of sperm DNA damage in infertile man could be useful to develop new therapeutic strategies and improve success of assisted reproduction techniques (ART).

scholarly journals Environmental Tobacco Smoke Exposure during Intrauterine Period, Promotes Caspase Dependent and Independent DNA Fragmentation in Sertoli-Germ Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Beril Yüksel ◽  
Sevtap Kilic ◽  
Nese Lortlar ◽  
Nicel Tasdemir ◽  
Semra Sertyel ◽  
...  
Keyword(s):  
Dna Damage ◽  
Cell Count ◽  
Cigarette Smoke ◽  
Sertoli Cell ◽  
Germ Cells ◽  
Cell Apoptosis ◽  
Smoke Exposure ◽  
Intrauterine Life ◽  
Group 2 ◽  
Group 1

Objectives. To investigate the effect of cigarette smoke exposure during intrauterine period on neonatal rat testis. Methods. Twenty-five rats were randomized to be exposed to cigarette smoke with the Walton Smoking Machine or to room air during their pregnancies. The newborn male rats (n=21) were grouped as group 1 (n=15) which were exposed to cigarette smoke during intrauterine life and group 2 (n=6) which were exposed to room air during intrauterine life. The orchiectomy materials were analyzed with TUNEL immunofluorescent staining for detection of DNA damage. To detect apoptosis, immunohistochemical analyses with caspase-3 were performed. Primary outcomes were apoptotic index and immunohistochemical scores (HSCORES); secondary outcomes were Sertoli-cell count and birth-weight of rats. Results. Sertoli cell apoptosis was increased in group 1 (HSCORE =210.6±41.9) when compared to group 2 (HSCORE =100.0±17.8) (P=0.001). Sertoli cell count was decreased in group 1 (P=0.043). The HSCORE for the germ cells was calculated as 214.0±46.2 in group 1 and 93.3±10.3 in group 2 (P=0.001) referring to an increased germ cell apoptosis in group 1. The apoptotic indexes for group 1 were 49.6±9.57 and 29.98±2.34 for group 2 (P=0.001). The immunofluorescent technique demonstrated increased DNA damage in seminiferous epithelium in group 1. Conclusions. Intrauterine exposure to cigarette smoke adversely affects neonatal testicular structuring and diminishes testicular reserve.

Association between germline DNA damage repair-related genes mutation and tumor mutational burden in lung cancer patients.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e21163-e21163
Author(s):  
Wei Nie ◽  
Hua Zhong ◽  
Ding Zhang ◽  
Shiqing Chen ◽  
Baohui Han
Keyword(s):  
Dna Damage ◽  
Dna Damage Repair ◽  
Gene Mutations ◽  
Related Gene ◽  
Mutational Burden ◽  
Damage Repair ◽  
Tumor Mutational Burden ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

e21163 Background: Deleterious somatic DNA damage repair (DDR) gene mutations are frequent in non-small cell lung cancer (NSCLC) and are associated with improved clinical outcomes of immunotherapy. DDR gene mutations are associated with higher tumor mutational burden (TMB) in cancer. However, the effect of germline DDR-related genes mutation with different functional annotations on TMB in NSCLC patients is still unclear. Methods: 1671 Chinese patients with NSCLC were enrolled in this study. Genomic profiling was performed on formalin-fixed paraffin-embedded tumor samples or peripheral blood by next generation sequencing (NGS) with 733 cancer-related genes panel. The germline mutation data were obtained. All annotations in clinical significance were according to the 2015 American College of Medical Genetics and Genomics-Association for Molecular Pathology (ACMG-AMP) guidelines. Results: 1076 patients (64.39%) had germline DDR-related gene mutations and 595 (35.61%) had no germline DDR-related gene mutations. Among patients with DDR-related gene mutations, 78 (7.25%) patients had the pathogenic (P) mutations or likely pathogenic (LP) mutations and 1056 (98.14%) had variants of unknown significance (VOUS) mutations. In total, the median TMB was 3.91 mutations/MB (range, 0-68.16) and 4.47 mutations/MB (range, 0-51.40) in patients with P, LP or VOUS mutations and no germline DDR-related gene mutations, respectively. To the further analysis, we divided patients with germline DDR-related gene mutations into three groups: only P or LP mutations (Group 1), only VOUS mutations (Group 2) and concurrence with P/LP/VOUS mutations (Group 3). Compare to the DDR-negative group, TMB was significantly lower in Group 2 (P < 0.001). No significant differences in Group 1 and Group 3 were observed. In addition, we found that mutations in different DDR pathway could not affect TMB value significantly. Conclusions: Germline DNA damage repair-related genes mutation may be not associated with TMB.

Poor sperm quality and advancing age are associated with increased sperm DNA damage in infertile men

Andrologia ◽  
2011 ◽  
Vol 44 ◽  
pp. 642-649 ◽  
Author(s):  
J. Varshini ◽  
B. S. Srinag ◽  
G. Kalthur ◽  
H. Krishnamurthy ◽  
P. Kumar ◽  
...  
Keyword(s):  
Dna Damage ◽  
Sperm Quality ◽  
Sperm Dna Damage ◽  
Infertile Men ◽  
Sperm Dna

scholarly journals A Cytogenetic Study on the Efficacy of Chyawanprash Awaleha as an Antioxidant in Oral Premalignant Cancer

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
A. N. Uma ◽  
Dhananjay S. Kotasthane
Keyword(s):  
Dna Damage ◽  
Precancerous Lesions ◽  
Cytogenetic Study ◽  
Mean Difference ◽  
Betel Quid ◽  
Satellite Association ◽  
Betel Quid Chewing ◽  
Group 2 ◽  
Oral Precancerous Lesions ◽  
Group 1

Background. Chyawanprash awaleha (Cp) is an Ayurvedic rasayana formulation and is used as a genoprotective agent. Objective. The present cytogenetic study has been done to investigate the efficacy of Cp against betel quid chewers suffering from oral precancerous lesions through satellite association (SA) assay. Materials and Methods. The frequency of SA was analyzed in 21 betel quid chewing oral precancerous lesions patients and then they were divided into 2 groups. Group 1 consisted of 15 patients, advised to quit betel quid chewing and fed with 20 gms of Cp, twice a day for three months. Group 2 consisted of 6 patients, who refused Cp feed but accepted to quit betel quid chewing. At the end of three months, both groups were assessed cytogenetically. Results. The frequency of SA was statistically significant in both groups, but an elevated mean difference was observed more in Group 1 than in Group 2. Conclusion. The study indicates that betel quid cessation reduces the effect of DNA damage in oral precancerous lesions. But the increased mean difference in SA in Group 1 compared to Group 2 clearly indicates that Cp can further minimize the genotoxic effect caused by mutagenic agents present in betel quid.

scholarly journals Rapid rates of sperm DNA damage after activation in tench (Tinca tinca: Teleostei, Cyprinidae) measured using a sperm chromatin dispersion test

Reproduction ◽  
2009 ◽  
Vol 138 (2) ◽  
pp. 257-266 ◽  
Author(s):  
Carmen López-Fernández ◽  
Matthew J G Gage ◽  
Francisca Arroyo ◽  
Altea Gosálbez ◽  
Ana M Larrán ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Dna Fragmentation ◽  
Living Organism ◽  
Sperm Chromatin ◽  
Tinca Tinca ◽  
External Fertilization ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Dispersion Test

Spermatozoal haplotypic DNA is prone to damage, leading to male fertility problems. So far, the assessment of sperm DNA breakage has been challenging because protamines render the nuclear chromatin highly compacted. Here, we report the application of a new test to quantify DNA fragmentation in spermatozoa of an externally fertilizing teleost fish. The sperm chromatin dispersion (SCD) test uses a species-specific lysing solution to generate controlled protein depletion that, followed by DNA-specific fluorescent labelling, allows an easy morphological discrimination between nuclei affected by DNA damage. Using tench (Tinca tinca) as our model, we first trialled the test against established, but more technically demanding, assays employing in situ nick translation (ISNT) and the comet assay. The SCD test showed high concordance with ISNT, comet assay measures and a chromatin-swelling test, confirming the application of this straightforward SCD technique to various aspects of reproductive biology. Second, we examined between-male variation in DNA damage, and measured changes through time following spermatozoal activation. Between-male variation in the basal levels of average DNA damage ranged from 0 to 20% of sperm showing damage, and all showed increases in DNA fragmentation through time (0–60 min). The rates of DNA damage increase are the fastest so far recorded in sperm for a living organism, and may relate to the external fertilization mode. Our findings have relevance for broodstock selection and optimizing IVF protocols routinely used in modern aquaculture.

scholarly journals The Effects of Alkaloids Fraction of Cyperus esculentus on Some Biochemical Parameters and Histopathology of the Testis of Lead-Induced Toxicity

2021 ◽  
pp. 55-62
Author(s):  
Godson Emeka Anyanwu ◽  
Luqman Adepoju Hassan ◽  
Ifeanacho Ezeteonu Abireh ◽  
Nto Johnson Nto
Keyword(s):  
Protective Effect ◽  
Normal Saline ◽  
Sperm Quality ◽  
Serum Testosterone ◽  
Wistar Rat ◽  
Saline Group ◽  
Cyperus Esculentus ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Aim: This study evaluated the protective effect of alkaloids fraction of Cyperus esculentus on lead-induced testicular toxicity in Wistar rat. Methodology: Twenty-five adult male Wistar rats were randomly divided into five (5) groups, (n=5). Group 1 was administered with 1ml normal saline only, group 2 was administered with 30 mg/kg of lead, group 3 was administered with 50mg/kg of alkaloids and 30 mg/kg of lead, group 4 was administered with 100 mg/kg of alkaloids and 30 mg/kg of lead, group 5 was administered 150mg/kg of alkaloids and 30 mg/kg of lead orally for 28 days. The testes of the rats were harvested on day 29 of the experiment and histological studies done using the H&E and Verhoeff-Van Gieson (VVG) stains. Sperm parameters, sex hormones and antioxidant of testicular homogenates were analysed. Results: Histological examination of the testes revealed increased spermatogenic cells and Leydig cell proliferation in the rats in groups 3, 4, and 5 administered with 50 mg/kg, 100 mg/kg, and 150 mg/kg of Alkaloids, and 30 mg/kg of lead, respectively, when compared with group 2 administered with 30 mg/kg of lead alone. Also there was significant increase in levels of serum testosterone (p < .05) in groups 3, 4, and 5 when compared with group 2. There was increase in levels of follicle stimulating hormone in groups 4, and 5 when compared with group 2. However, significant decreased in luteinizing hormone was observed in groups 3, 4 and 5. Group 2, treated with 30 mg/kg of lead only showed increased malondialdehyde levels when compared with group 1 that received 1ml normal saline. Group 3, 4, and 5, treated with 50 mg/kg, 100 mg/kg, and 150 mg/kg of alkaloid plus 30 mg/kg lead, respectively, showed significant growth of seminiferous epithelium, improved sperm quality, and decreased levels of malondialdehyde (p < .05). Conclusion: This study shows that alkaloids fraction of Cyperus esculentus may have a protective effect on the testis of Wistar rat when it is exposed to toxicity from lead.

113 EFFECT OF SPERM COATING ON THE QUALITY OF BOVINE FROZEN - THAWED SPERMATOZOA

2006 ◽  
Vol 18 (2) ◽  
pp. 164
Author(s):  
M. Thys ◽  
A. Van Soom ◽  
J. Dewulf ◽  
T. Rijsselaere ◽  
A. de Kruif
Keyword(s):  
Seminal Plasma ◽  
Sperm Quality ◽  
Univariate Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Freezing And Thawing ◽  
Bovine Sperm ◽  
Sperm Characteristics ◽  
Group 2 ◽  
Group 1

The substantial decrease of sperm quality after cryopreservation remains an important issue in the artificial insemination industry. Sperm coating with Triladyl® (Minitübe, Tiefenbach, Germany) during ejaculation can preserve sperm characteristics and oocyte penetrating capacity of fresh bovine spermatozoa stored in egg yolk diluent for up to 6 days (De Pauw et al. 2003 Theriogenology 59, 1109–1122). Since collecting semen in a tube containing egg yolk-Tris extender (sperm coating) limits the period of contact between spermatozoa and seminal plasma, the present experiment was conducted to assess if this slightly adjusted method of sperm collection could also have a significant effect on bovine sperm quality after cryopreservation. Semen of five young Holstein Friesian bulls was collected by means of an artificial vagina connected to an empty tube (Group 1; five ejaculates per bull) or a tube containing 4 mL of an egg yolk-Tris extender (Groups 2 and 3; each five ejaculates per bull). The semen samples of Group 1 were conventionally diluted in straws (60 × 106 sperm/mL), frozen, and stored in liquid nitrogen. The samples of Group 3 were centrifuged, and after removing diluent and seminal plasma, the sperm pellet was conventionally diluted and processed. The samples of Group 2 were processed without removal of the supernatant. After thawing each ejaculate was analyzed for average path velocity (VAP), beat cross frequency (BCF), and progressive motility (PROG) using CASA (Minitübe, Tiefenbach, Germany). Furthermore, the membrane integrity of each sample was evaluated using fluorescent SYBR®–14/PI staining (BD Biosciences, Erembodegem, Belgium). All parameters were compared among the three groups of sperm using univariate analysis of variance (SPSS 12.0; SPSS, Inc., Chicago, IL, USA). No significant differences could be observed among the three groups for all of the evaluated sperm characteristics (Table 1). A significant effect of the bull could be determined for all analyzed parameters (P ≤ 0.02), except for the percentage of moribund cells. Nevertheless, the group-bull interaction was never statistically significant. Coating bovine sperm with an egg yolk-Tris extender during ejaculation cannot prevent the substantial deterioration of the spermatozoa that occurs during freezing and thawing since this method of sperm collection does not significantly influence the motility parameters or the membrane integrity after thawing. Table 1. VAP, BCF, PROG, and percentage of membrane-intact, dead, and moribund spermatozoa for the three groups of sperm This research was supported by IWT (no. IWT/020727).

68 DIFFERENCE IN THAWING CURVE OF STALLION FROZEN SEMEN DILUTED IN 2 EXTENDERS

2015 ◽  
Vol 27 (1) ◽  
pp. 127
Author(s):  
C. P. Freitas-Dell'aqua ◽  
C. Ramires Neto ◽  
Y. F. R. Sancler-Silva ◽  
P. M. Papa ◽  
J. A. Dell'aqua ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Freezing Process ◽  
Freezing And Thawing ◽  
Becton Dickinson ◽  
Plasma Membrane Integrity ◽  
Frozen Semen ◽  
Group 2 ◽  
Artificial Vagina

Commercial freeze extenders have different composition and ratio of cryoprotectors; freezing and thawing protocols are different for each extender. The aim of this experiment was to observe the effect of thawing curve in stallion frozen semen with 2 commercial extenders. Two ejaculates from each of 9 stallions of different breeds (Quarter Horses and Mangalarga Marchador) were used. Semen was collected using an artificial vagina, and the ejaculate was divided into 2 groups following the manufacture's protocol: group 1 (INRA), in which the semen was diluted 1 : 1 with the extender INRA 96TM (IMV, Paillette Crista, France) and group 2 (BC), in which the semen was diluted (1 : 1) with the extender Botu-SemenTM (Botupharma, Brazil). The samples of the 2 groups were centrifuged at 600 × g for 10 min, the supernatant was discarded, and the pellet was resuspended with INRA FreezeTM (group INRA, IMV) and with BotucrioTM (group BC, Botupharma) at the concentration of sperm 100 × 106 sperm mL–1. After this, the semen was packaged in 0.5-mL straws. For each group the freezing process was carried out according to the manufacturer's instructions. The straws were thawed in a water bath with 3 different thawing curves: 37°C for 30 s (37/30), 46°C for 20 s (46/20), and 75°C for 7 s (75/7) before analysis. The aim of these rates is to keep the semen in 37°C post-thaw. The sperm kinetic analysis was performed by computerized method (CASA, HTM-IVOS, IMV, USA) and the analysis of plasma membrane integrity by flow cytometer (BD LSR Fortessa, Becton Dickinson, Mountain View, CA, USA). Data of sperm kinetic and of plasma membrane integrity were compared among the 3 thawing curves for one extender using analysis of variance. Differences were considered significant at a probability level of 5%. No differences were observed in total motility (%, BC 37/30 = 72.8 ± 14.4; BC 46/20 = 70.0 ± 14.2; BC 75/7 = 70.3 ± 12.0 v. INRA 37/30 = 57.2 ± 19.1; INRA 46/20 = 50.0 ± 21.9; BC 75/7 = 58.8 ± 20.8), progressive motility (%, BC 37/30 = 36.9 ± 8.2; BC 46/20 = 34.4 ± 10.5; BC 75/7 = 33.6 ± 7.8 v. INRA 37/30 = 25.3 ± 12.7; INRA 46/20 = 21.9 ± 13.9; BC 75/7 = 28.9 ± 14.8), rapid sperm (%, BC 37/30 = 59.7 ± 16.4; BC 46/20 = 56.8 ± 17.1; BC 75/7 = 58.1 ± 14.9 v. INRA 37/30 = 38.3 ± 20.9; INRA 46/20 = 35.3 ± 22.9; BC 75/7 = 44.4 ± 23.8), and plasma membrane integrity (%, BC 37/30 = 49.1 ± 14.8; BC 46/20 = 43.1 ± 13.1; BC 75/7 = 46.7 ± 11.8 v. INRA 37/30 = 32.2 ± 10.7; INRA 46/20 = 29.6 ± 10.1; BC 75/7 = 37.4 ± 9.1) among the 3 thawing curves for INRA and BC groups. In this study, we can conclude there is no influence of the 3 tested thawing curves in sperm quality for stallion frozen semen with INRA Freeze and Botucrio extenders.

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