Effect of sperm DNA fragmentation on blastocyst formation rate and subsequent IVF outcome

Background: Male infertility associated with sperm DNA alteration has raised a new issue in assisted reproduction techniques (ARTs).Methods: It was a retrospective analytical study on 250 cases of routine IVF/ICSI performed at Swagat ART Centre from January 2017 to January 2020. We divided the patient according to the sperm DNA fragmentation index (DFI) as normal DFI≤15%, n=95, a moderate DFI≤30%, n=89, and a high DFI group >30%, n=66. Oocytes of each patient were almost equally divided and fertilization method was adopted as half IVF half ICSI or only ICSI in poor quality (oligo, astheno, teratozoospermia or with two or all three defect and compared the fertilization, cleavage, embryo formation, blastocyst formation, pregnancy and early embryo formation rate among these six groups. Results: Fertilization, cleavage, embryo formation, and clinical pregnancy rates were reported as higher in ≤15% DFI group of both IVF and ICSI-ET (87.3±26.2, 77.7±26.1, 68.2±28.8, 50.8 in IVF and 78.3±17.8, 70.3±31.2, 67.2±28.8, 57.6 respectively). Significant differences (p<0.01) are observed among all six groups. Higher abortion rate is observed in high DFI group of both IVF and ICSI.Conclusions: High sperm DFI causes low blastocyst formation and pregnancy outcome. Higher abortion rate observed in high DFI group indicated need of further study.

Assessing Genotoxicity of Ten Different Engineered Nanomaterials by the Novel Semi-Automated FADU Assay and the Alkaline Comet Assay

Increased engineered nanomaterial (ENM) production and incorporation in consumer and biomedical products has raised concerns about the potential adverse effects. The DNA damaging capacity is of particular importance since damaged genetic material can lead to carcinogenesis. Consequently, reliable and robust in vitro studies assessing ENM genotoxicity are of great value. We utilized two complementary assays based on different measurement principles: (1) comet assay and (2) FADU (fluorimetric detection of alkaline DNA unwinding) assay. Assessing cell viability ruled out false-positive results due to DNA fragmentation during cell death. Potential structure–activity relationships of 10 ENMs were investigated: three silica nanoparticles (SiO2-NP) with varying degrees of porosity, titanium dioxide (TiO2-NP), polystyrene (PS-NP), zinc oxide (ZnO-NP), gold (Au-NP), graphene oxide (GO) and two multi-walled carbon nanotubes (MWNT). SiO2-NPs, TiO2-NP and GO were neither cytotoxic nor genotoxic to Jurkat E6-I cells. Quantitative interference corrections derived from GO results can make the FADU assay a promising screening tool for a variety of ENMs. MWNT merely induced cytotoxicity, while dose- and time-dependent cytotoxicity of PS-NP was accompanied by DNA fragmentation. Hence, PS-NP served to benchmark threshold levels of cytotoxicity at which DNA fragmentation was expected. Considering all controls revealed the true genotoxicity for Au-NP and ZnO-NP at early time points.

Novel coumarin derivatives containing a triazole moiety: A study on synthesis, cytotoxicity, membrane dysfunction, apoptosis, cell cycle, and antiangiogenic studies

Background: Coumarin is a functional compound with a pronounced wide range of biological activities and has recently been shown to have anticancer effects on various human cancer cells. Cisplatin is widely used in treating many cancers, but its effectiveness is limited due to acquired resistance and dose-related side effects. Objective: This study aimed to reveal the chemosensitizing ability of novel synthesized coumarin-triazole hybrid compounds (3a-f) compared to the cisplatin in A549, MCF-7, and HeLa cancer cells. Methods: Cytotoxicity was determined by MTT assay. Lactate dehydrogenase (LDH), antioxidant/oxidant status, DNA fragmentation were determined spectrophotometrically using commercial kits. Muse™ Cell Analyzer was used to assess cell cycle progression. Pro/anti-apoptotic gene expressions were determined by Real-Time qPCR. The antiangiogenic activity was determined by VEGF expression and Hen's chorioallantoic membrane model. Results: Compounds 3c, -d, -e, and -f potentiated the cisplatin-induced cytotoxicity through the increased LDH release and DNA fragmentation, induced G2/M cell cycle arrest, overproduction of oxidative stress, and decrease of cellular antioxidant levels. These compounds combined with cisplatin caused upregulation in the pro-apoptotic Bax, Bıd, caspase-3, caspase-8, caspase-9, Fas, and p53 gene expressions while downregulating anti-apoptotic DFFA, NFkB1, and Bcl2 gene expressions. These combinations caused vascular loss and a reduction in VEGF expression. Conclusion: These results suggest that a combinational regimen of coumarin compounds with cisplatin could be enhancing the effect of cisplatin in A549 cells. Besides, considering compounds have relatively low toxicity in normal cells, they decrease the dose requirement of cisplatin in cancer treatments.

Relationship Among Traditional Semen Parameters, Sperm DNA Fragmentation, and Unexplained Recurrent Miscarriage: A Systematic Review and Meta-Analysis

Several studies have explored the relationship among traditional semen parameters, sperm DNA fragmentation (SDF), and unexplained recurrent miscarriage (RM); however, the findings remain controversial. Hence, we conducted a meta-analysis to explore the relationship among traditional semen parameters, SDF, and unexplained RM. Multiple databases, including PubMed, Google Scholar, MEDLINE, Embase, Cochrane Library, Web of Science, and China National Knowledge Infrastructure (CNKI), were searched to identify relevant publications. From the eligible publications, data were extracted independently by two researchers. A total of 280 publications were identified using the search strategy. According to the inclusion/exclusion criteria, 19 publications were eligible. A total of 1182 couples with unexplained RM and 1231 couples without RM were included in this meta-analysis to assess the relationship among traditional semen parameters, SDF, and unexplained RM. Our results showed that couples with unexplained RM had significantly increased levels of SDF and significantly decreased levels of total motility and progressive motility compared with couples without RM, although significant differences were not observed in the semen volume, sperm concentration, and total sperm count between couples with and without RM. The SDF assay may be considered for inclusion in evaluations of couples with unexplained RM.

Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos

This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio) embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified (Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 µM (T2) and 1 mM of melatonin (T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos stored in -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2) and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentation analyses were performed. No live embryos were obtained from vitrified treatments, observing a rapid degeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followed by breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 had the highest relative gene expression, for the three genes (P < 0.05) furthermore, T2 had similar expression of pro-apoptotic genes to CG (P < 0.05). ROS formation revealed that CG presented lower percentage of embryo surface area affected (3.80 ± 0.40%) (P < 0.05), in contrast, no differences were found among the other groups. T1 was most significantly (P < 0.05) damaged by DNA fragmentation. The vitrified groups with melatonin had similar damage levels of CG (P > 0.05). The inclusion of 1 µM of melatonin in the vitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utility in cryopreserving fish embryos.

Bubble-enhanced ultrasonic microfluidic chip for rapid DNA fragmentation

We present a new bubble-enhanced microfluidic approach for highly efficient DNA fragmentation, suitable for next generation sequencing platforms. Improved on-chip performance arises from acoustic streaming generated by oscillating bubble interfaces.

The effect of preparatory therapy with myo-inositol on the outcome of infertility treatment by ART in men with low reproductive function

Research objective: to study the effect of myo-inositol (MI), folic acid (FA) and cyanocobalamin in mono- and complex therapy on the classical fertility rates and the level of DNA fragmentation in sperm in men with low reproductive function, and on the results of the in vitro fertilization (IVF) programs performed in these patients.Materials and methods. A retrospective study of 133 patients aged 32 to 40 years was carried out for the period 2020–2021. All patients were divided into three groups: group 1 – 44 men who took MI, FA and cyanocobalamin daily as monotherapy for 3 months; group 2 – 42 patients who followed a diet to improve spermatogenesis for 3 months; group 3 – 47 men, for whom complex therapy was applied for 3 months: MI, FA, cyanocobalamin and diet.Results. The increase of sperm concentration in ejaculate, improvement of progressively motility of spermatozoa, and increase of the percent of spermatozoa with normal morphology in men with failures of reproductive function after the complex preparatory therapy are defined. The preparatory therapy for men with MI in preparation for IVF helps to improve the quality of embryos and increase the pregnancy rates in couples with male infertility due to the assisted reproductive technologies. The positive effect of preparatory complex therapy in patients with poor spermatogenesis on the blastocyst formation rates in IVF programs is shown.Conclusions. The positive effect of MI on the normal functioning of the reproductive system in men has been proven. The increase pregnancy rates in the IVF program after preparatory treatment of men with MI can be explained by its positive effect on reducing the level of DNA fragmentation of sperm.