Background: Varicocele is an abnormal dilation of the testicular veins in the pampiniform plexus and characterized by retrograde blood flow in the internal spermatic veins due to incompetent or absent valves. It is considered one of the main causes of male factor infertility. Sperm DNA integrity is essential for in vivo and in vitro fertilization, and varicocele causes a progressive decrease in sperm quality . Objective: To study the impact of varicocele on semen quality and sperm DNA integrity. Patients, Materials and Methods: Twenty patients with varicocele and 20 men without varicocele were assigned into two groups in the Out-Patient Clinic of Urology in Al-Kadhimya Teaching Hospital . Semen was collected from both groups and the rate of sperm DNA fragmentation assessed by the "Comet assay" and categorized to 4 classes. Class I (no DNA fragmentation), class II (little DNA fragmentation), class III (meaningful DNA fragmentation), and class IV (high DNA fragmentation) . Results: No statistically significant differences were found between the varicocele and control groups with respect to age. Men with varicocele had significantly higher ejaculate volume, lower sperm concentration, and lower progressive motility, and showed more abnormal morphology than men in the control group. The Varicocele group showed a lower percentage of sperm with little DNA fragmentation (class II) and a higher percentage of sperm with DNA fragmentation (class IV), than the control group. Conclusions: Compared with men without varicocele, men with varicocele had a higher percentage of cells with DNA fragmentation .
One-half of the genome is delivered by the sperm, and a healthy sperm is needed for fertilization, fetal development, and child wellbeing. Abnormal sperm DNA can impact any of these processes. The measurement of sperm DNA fragmentation has been added to some medical andrology laboratories as a crucial addition to the standard analysis of the semen. Sperm DNA fragmentation has also been linked to male infertility including idiopathic male infertility. Some degree of controlled DNA nicking is essential for adequate DNA compaction, but excessive sperm DNA fragmentation is usually associated with reduced male fertility potential, reduced fertilization, poor embryo quality, recurrent pregnancy loss and poor assisted reproductive techniques outcomes. Semen samples from 50 infertile males (as 19 asthenozoospermia, 7 oligozoospermia, 13 normospermia and 11 combined defects) were analyzed for sperm DNA fragmentation. This study followed the World Health Organization 2010 guidelines to assess the sperm parameters. Sample were collected by masturbation and put in a 37 °C incubator to be liquefied, then they were placed in a centrifuge (3000 rpm for 10 min) to extract the pellet, which was used for assessment of DFI% by anillin blue staining. Negative significant correlation (p = 0.023) between morphologically normal sperms and DFI % was found in normozoospermia group. The objective of this study is to examine the correlation of DFI among different seminal fluid categories (normozoospermia, oligozoospermia, asthenozoospermia and, combined).
When evaluating male reproductive potential, it is crucial to evaluate sperm structural abnormalities and sperm DNA damage. Apoptosis and mitochondrial membrane potential disruption are linked to sperm DNA fragmentation (SDF), which is inversely correlated with motility and normal morphology. To enable liquefaction, the semen sample is taken in a sterile collection container and incubated at 37°C for 30 minutes The samples were then split into three groups, P, G, and N, based on the morphological %. To measure chromatin condensation, aniline blue staining (AB), an acidic dye with a strong affinity for non-lysine-rich histones, was employed. The DNA maturity of the spermatozoa was assessed using AB. Sperm concentration, total sperm count, total motility, and total progressively motile sperm all had means that were above the WHO 2010 standard range. The DNA fragmentation index was significantly higher in patients with pattern P than in patients with patterns G and N, and it differed significantly (P=0.003) between the semen morphological patterns. In conclusion, as DNA fragmentation is an independent test this study showed that a low percentage of normal sperm morphology might indicate DNA integrity status.
Male infertility is a global health issue affecting over 20 million males. A spermatogenesis-related cell differentiation abnormality causes teratozoospermia (TZS). Extensive research has linked TZS to poor fertilization outcomes at assisted reproductive technology (ART) centers as well as to spontaneous pregnancies. This study used Glycyrrhiza glabra (Gg) and Platelet Rich Plasma (PRP) media to establish a culture medium that aimed to improve certain sperm functions of teratozoospermic men in vitro. A total of fifty semen samples were examined. The World Health Organization 2021 and 1999 standards were used to evaluate certain sperm parameters and the DNA fragmentation index. Platelet-rich plasma and Gg extract were utilized as a medium for in vitro preparation and activation of semen samples complaining of teratozoospermia factor. Following activation by the utilization of the density gradient approach and the combination of Gg and PRP medium, there was a statistically significant ( p <0.001) improvement in certain sperm function metrics. These improvements included a higher percentage of morphologically normal sperm, a rise in active motility, and a drop in the DNA fragmentation index. The density gradient technique, regardless of the medium used in this investigation, resulted in a significant enhancement of morphologically normal sperms with other certain sperm parameters and a decrease in the DNA fragmentation index.
For conception and the development of healthy embryos, sperm DNA integrity is crucial. According to a growing body of studies, there is a strong correlation between sperm DNA damage and male infertility. Among the new medicines being developed in the medical field, the application of Platelet Rich Plasma (PRP) in human reproduction has yet to be examined. A total of 100 semen samples were used in the current experimental investigation. From November 2020 to June 2021, the research was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Masturbation was used to get an ejaculated semen sample. After semen analysis, the samples were separated into two equal parts, one without autologous PRP and the other with 2% autologous PRP, with the DNA fragmentation assessed using the Sperm Chromatin Dispersion Test. There was highly significant reduction in DNA fragmentation index (p < 0.001). The mean sperm DNA integrity was reduced after adding PRP (33.85±16.73 vs 38.55±16.64), Mean (± SE). PRP has been shown to improve human sperm DNA integrity.
Background: Semen cryopreservation is a useful tool for preserving fertility for men involved in ART program who have been diagnosed with cancer and will undergo chemotherapy, radiotherapy or testicular surgery. Objective: The objective of this study to evaluate the effect of honey bee supplemented to cryoprotectant medium on post-thaw sperm DNA integrity ,morphology and round cells. Subjects ,Materials and Methods: Thirty semen samples were collected from 30 infertile patients. After assessment of semen analysis, semen samples were divided into 3 aliquots (0.7ml for each sample) and mixed with 1 ml of cryopreservation solution (G1,control) alone, enriched with 5% honey bee(G2) with 10% honey bee (G3) for cryopreservation. Current study used the acridine orange test to investigate the effect of honey bee in two concentration (5%, 10%) on the integrity of sperm chromatin structure pre-and post cryopreservation. Cryopreservation was done at -196 Co in liquid nitrogen and thawing was performed after six months. Direct swim up technique was used for in vitro sperm preparation post-thawing, sperm parameters were assessed and data were statistically analyzed pre-and post-thawing. Results: The results show there was a significant(P<0.05) increase in the percentage of morphologically normal sperm for all groups post-thawing, particularly for G3 group that was significantly (P<0.05) increased as compared to G1 and G2 groups, In contrast, non-significant differences (P>0.05) were observed between G1 and G2 groups. Sperm DNA fragmentation percentage (%) was significantly(P<0.05) increased in G1 post thawing without any additives as compared to pre-cryopreservation , where as in G2 and G3 sperm DNA fragmentation were increased but not significantly as post-cryopreservation groups. Also the results show that there were no significant differences between G2 and G3 whereas G3 gives better result as compared to G1 with significant differences. Round cells counts for all groups of post cryopreservation were significantly(P>0.05) decreased as compared to pre-cryopreservation. Non significant (P>0.05) differences in the round cells counts among all the groups of post -thawing in spite of G3 group has the lowest mean. Conclusions: From these results we concluded that supplementation of 10% of honey bee to the freezing cryoprotectant medium gives best protection to the morphology and DNA integrity of human sperm.
Background: Citrullus colocynthis (CC) is an herbal medicine used in the treatment of a wide range of diseases. Recently, It has been studied on in vitro activation of mice sperms with no research concern the effect of this plant on the DNA normality following in vitro preparation of human semen samples. Objectives: The present study was aimed to examine the effect of adding CC in the medium used for in vitro sperm activation of asthenospermic men and related factors and to found out the effect of CC on the DNA status following the activation. Materials and Methods: Citrullus colocynthis aqueous extract (0.01mg/ml culture medium) was used for in vitro sperm activation . One hundred and ten semen samples were divided into two groups, the first group activated in vitro by adding CC extract to the culture medium Ham's F-12 (the treated group) and, the second group sperm activated in vitro with Ham's F-12 alone ( the control group). Results: In vitro activation of sperms with aqueous extract of CC has shown positive effect on sperm concentration, sperm motility, and grade activity of progressive forward movement. There was a significant (P<0.05) increase in certain sperm function parameters of asthenozoospermic in treated group compared with the control group .The study showed that the DNA fragmentation resulted from treated group were lower than that of control group. Conclusions: According to the results of present study, adding aqueous extract of CC to the human sperms activation culture medium leads to an improvement in certain sperm function parameters of asthenozoospermic men with no harmful effect on the human sperms DNA with decrease in DNA denaturation.
Cigarette smoke contains a substance which is a toxic mutagenic chemical, and carcinogenic compounds, which have a negative effect on male fertility. Smoking cigarettes is associated with noticeably higher levels of the oxidative stress-causing reactive oxygen species in the seminal fluid. Because of this, smoking may cause changes to the sperm plasma membrane and a significant amount of deoxyribonucleic acid fragmentation. To study the influence of smoking on sperm chromatin immaturity and ICSI outcomes (fertilization rate, embryo grading and pregnancy rate). At the laboratories of the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies and after 1-7 days of sexual abstinence, 92 males’ sperm samples were taken. After liquefaction, macro-and microscopic studies were carried out, criteria applied by the WHO 2021. Masturbation was used to obtain the sample, which was then placed in sterile containers. The relationship between the smoking and chromatin immaturity and ICSI outcomes have been evaluated. A significant positive correlation between smoking status and sperm chromatin immaturity. Furthermore, there was a negative non-significant correlation between smoking status and fertilization rate. Weak and non-significant correlations between smoking status and embryo grade 2 as well as embryo grade 3. There was a positive association between smoking status and embryo grade 1. The sperm chromatin integrity was significantly lower in the non-smoker group compared to the smoker group.
Background: The immune infertility caused by anti-sperm antibodies (ASAs) represented about 1020%- of infertility among the couples, the ASAs interfere with sperm parameters such as the sperm motility and sperm ability to penetrate cervical mucus, sperm-oocyte binding, and fertilization and embryo developments. Objectives: The present study designed to assess semen analysis, presence of ASAs and DNA fragmentation index as well as correlation within these parameters in normzoospermic Iraqi subjects Patients, Materials and Methods: A total number of Iraqi subjects (116) with range of age (2051-) years and their mean duration of infertility (4.70 ± 2.77). Seminal fluid for Macroscopic and Microscopic assessments done according to WHO 2010 criteria. The mixed agglutination reaction (MAR) test used to assess theirASAs in semen (direct method), seminal plasma and serum (indirect method); for the both IgG and IgA classes of antibodies and their distribution among different parts of spermatozoa as percentage. Results: The mean of IgG using direct method was (23.881.75±) and for indirect detection method the mean was (27.412.41±) with no significant difference (P<0.05) between the two methods. While the direct method for IgA detection mean (14.46 ±1.76) and the mean for indirect method of detection (6.860.39±) and there was significant difference (P>0.05) between the two methods. In addition to that, the distribution of IgG and IgA detected by direct method on sperm parts showed no significance difference except for the sperm tail; while the indirect method for IgA and IgG showed significant difference in distribution on sperm mid-piece. Conclusion: The correlation among sperm parameters and Immunoglobulin using direct and indirect methods showing variations in significance also in the correlation between the direct and indirect methods according to distributions on sperm surface
The integrity of sperm deoxyribonucleic acid (DNA) and chromatin is very important for genetic material transmission into offspring. Therefore, the aim of the present study was to investigate the correlation between sperm deoxyribonucleic acid integrity and sperm parameters. The current study was conducted on 96 semen samples. This study was done at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, for the period between November 2020 and May 2021. The samples were collected, and seminal fluid analysis was performed according to World Health Organization guidelines. Sperm Chromatin Dispersion test was performed. The results showed that the deoxyribonucleic acid fragmentation of sperms in infertile men in this study was higher than reported in fertile men in previous studies, indicating deoxyribonucleic acid fragmentation (DNA) is actively contributing at least partially to male infertility. In terms of enrolled men, sperm deoxyribonucleic acid fragmentation (SDF) showed a significant positive correlation with round cell count.
Human sperm chromatin and integrity of DNA have multiple endogenous factors, such as sperm compression and nucleoproteins, with various mechanisms all through their own sperm production and transport. The integrity of the paternal genetic material in both regular and assisted pregnancy is essential for initiating and maintaining a viable pregnancy. The study included semen samples from 100 men taken from Al Nahrain University Medical Clinic and Babylon Teaching Hospital for Women and Children. The first sperm test was based on the World Health Organization (1999), followed by a DNA integrity test that uses a DNA fragmentation test, and the third intensification test Chromatin by aniline blue dye. It was correlated negatively with the sperm count in combination with DFI and chromatin sperm with semen characteristics (r = - 0.223, p = 0.026). As well as DFI, sperm chromatin was being clinically significant associated with the rate of round cells (r = 0.39, < 0.001). DFI was positive and significantly associated with sperm chromatin in all infertile men (r = 0.958; p<0.001). DNA isolation in all enrolled men has been adversely linked with sperm/ejaculation, has been associated with the round cell count positive way and is not understanding and resolving with other parameters. The width of abstinence and beneficial cell count in all register men were strongly linked with Sperm chromatin. There was a direct relationship between the fragmentation of DNA and chromatin. This could imply that one exam can be used instead of the other.
Exposure to radiation of various forms is one of the many environmental, health, and lifestyle variables that have been linked in recent years to an increase in the rate of male infertility. According to data from several recently published studies, mobile phone use may negatively affect sperm parameters, which may lower male fertility. Ninety-seven samples of semen from fertile and infertile individuals were sent to the "High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Al-Nahrain University in Baghdad, between November 2022 and May 2023. The samples were split into four groups: normozoospermia (n = 40), asthenozoospermia (n = 25), oligozoospermia (n = 16), and teratozoospermia (n = 16). Each group was further divided into two groups based on the amount of time spent using a mobile phone each day: less than 4 hours per day and equal to or more than 4 hours per day. A routine seminal fluid analysis was carried out to measure the sperm parameters and test the deoxyribonucleic acid fragmentation rate using an acridine orange stain. Samples exposed to electromagnetic waves of cell phones showed a significant decrease in DNA normality. deoxyribonucleic acid fragmentation index (p < 0.001) in normozoospermia patients exposed to more than 4 hours/day of non-ionizing electromagnetic waves. The results of deoxyribonucleic acid fragmentation index after treatment were shown a significant improvement compared to before the treatment in all fertile and infertile groups. Electromagnetic waves emitted from cell phones are associated with a decrease of deoxyribonucleic acid normality when exposure for <4hours/day. The study recommend that men of reproductive age avoid keeping their cell phones in the hip pocket and front pants pocket which may negatively affect spermatozoa deoxyribonucleic acid integrity and impair male fertility.