Reactive oxygen species (ROS) are believed to play an important role in the function of the male and female reproductive system, as well as in embryonic development. If the balance between production and elimination of ROS is disturbed, pathogenic consequences would happen. Reactive oxygen species can sometimes exert damaging effects when overabundant, oxidative stress is hypothesized to have a negative impact on, for example, folliculogenesis, embryo development, oocyte maturation, oocyte fertilization, endometriosis, polycystic ovary syndrome and unexplained infertility .8-Hydroxy2-desoxy guanosine (8‐OHdG) is a sensitive indicator of DNA damage as a result of oxidative stress. The study aim was to evaluate follicular fluid levels of 8-Hydroxy2-desoxy guanosine in patients undergoing Intracytoplasmic Sperm Injection (ICSI) and compared the results between both pregnant and non-pregnant patients . One hundred and twenty-seven women included in this study, they underwent controlled ovarian hyper stimulation, program followed by ova pick up, at that time follicular fluid were obtained and freezes till the time of analysis for reactive oxygen species (ROS), patient injected oocyte was cultured in CO2 incubator, then embryo grading, embryo transfer, pregnancy rate and reactive oxygen species level were determined. Follicular fluid 8-OHdG was significantly lower in pregnant females (p < 0.001), 8-OHdG had negative correlations with grade 1 and grade 2 embryos and negative correlation with total blastocyst count. Follicular fluid levels of 8-OHdG provides sensitive tool to predict positive pregnancy outcome in women undergoing ICSI.
Reactive oxygen species (ROS) are active chemical compounds created by the body's metabolic process and are crucial in human reproduction. Excessive ROS can contribute to oxidative stress, which has been linked to poor fertility. Embryo culture medium plays an essential role in IVF/ICSI cycles. Eighty participants underwent ovarian stimulation and ICSI cycles; ROS levels were measured in blood samples on the ovum pickup (OPU) day and in each embryo culture dish on ET day using enzyme-linked immunosorbent assay (ELISA). These values were associated with the quality of generated embryos. Women were divided into two groups; the first had day three embryo transfers, whereas the second received blastocyst transfers on day 5. Grades I embryo count was shown to be inversely related to the serum and culture medium ROS levels. Pregnant women had significantly greater levels of GI embryos (p=0.011) and lower levels of GIII embryos (p=0.024). Pregnancy rates for the third-and fifth days of the embryo transfers were 24% and 50%, respectively, with (p=0.045). Pregnancy success rates were increased after ET day five and were strongly influenced by transfer day. Blastocyst-stage embryos with high serum ROS levels were found to be underdeveloped. Culture medium ROS levels can be used as an adjuvant criterion for embryo selection and metabolic marker for developing embryos.
Infertility is described as a couple's failure to conceive for at least a year, using unprotected sex. Even though all anesthetic chemicals have been revealed in the follicular fluids, general anesthesia is still used in many IVF centers for patients who want to get pregnant. This study included 60 infertile women randomly divided into two groups under general anesthesia. 30 patients were given a Ketamine dose of 0.5 mg/Kg and the other 30 patients were received Remifentanil dose of 0.5 µg/Kg for induction of anesthesia. We collected serum for Reactive Oxygen Species (ROS) levels assessment for all patients before and after starting general anesthesia. Routine ICSI procedures was performed on all participants, including clinical evaluation (history, examination, and investigation), controlled ovarian and ovulation stimulation, oocyte retrieval under general anesthesia, follicular fluid collection for postoperative anesthesia medication concentration (Remifentanil and Ketamine), oocyte stripping, oocyte maturity assessment, intra-oocyte sperm injection into mature cells (MII), fertilization and division evaluation and embryo categorization, embryo selection and transfer, luteal phase support, and beta-hCG determination. ROS concentrations were compared between the Remifentanil and Ketamine groups. There was no significant difference in embryo features between Remifentanil and Ketamine, indicating that neither one is superior to the other in this regard. When Remifentanil or Ketamine was taken, there was no significant difference in ROS levels in serum or follicular fluid.
Background: Chemotherapy and radiotherapy can destroy or severely reduce spermatogenesis and thereby jeopardize fertility in the long term. There is still no medical treatment that guarantees fertility preservation after chemo- and radiotherapy. Now with recent improvement of assisted reproductive technologies (ART) and possibility of using testicular spermatozoa or epididymal spermatozoa at in vitro fertilization (IVF) by intracytoplasmic sperm injection (ICSI), cryopreservation of testicular tissue is an option in fertility preservation for males who will lose spermatogenic cells as a result of chemoand radiotherapy and for males with azoospermia. Objectives: This study is an attempt to evaluate oxidative DNA damage in the mice testicular tissue after cryopreservation/thawing cycle when using different types of cryoprotectants as well as to investigate the changes that occur in mice testicular tissue after cryopreservation/thawing cycle as a model for human being. Methods: Fifty mature fertile male mice between 8- 12 weeks old were used in the current study. For each mouse, one testis was evaluated histologically and immunohistochemically (using 8-Hydroxy 2'-Deoxyguanosine as marker of oxidative DNA damage) without cryopreservation (control group). The other testis was evaluated histologically and immunohistochemically (using 8-Hydroxy 2'-Deoxyguanosine as marker of oxidative DNA damage) after six weeks of cryopreservation using different types of cryoprotectants (cryopreserved group). Results: The microscopically observation of slides obtained from cryopreserved testis differs according to the type of cryoprotectant (glycerol, 1,2 propanediol and dimethylsulfoxide), in the dimethylsulfoxide group, tissue sections showed no major differences versus control group, but in the histological sections obtained from tissue cryopreserved by glycerol as cryoprotectant showed moderate morphological and structural changes as compared with the control group. While, the tissue samples subjected to 1,2 propanediol as cryoprotectant displayed the severest morphological and structural changes as compared with the control group. Immunohistochemical results of the present study showed a highly significant (P<0.001) increase in oxidative DNA damage in the cryopreserved testis (46.78%, 63.45% and 32.59% represent cryoprotectant glycerol, 1, 2 propanediol and dimethylsulfoxide, respectively) compared with control group (19.27%) after six weeks of cryopreservation. Conclusion: From the results of the current study, it was concluded that there was alteration in testis histology after cryopreservation/thawing cycle and increase the level of oxidative DNA damage after cryopreservation/thawing cycle. As well as dimethylsulfoxide cryoprotectant provide good protection for testis histology and DNA.
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.
Human peptidoglycan recognition protein-1 (PGLYRP‐1) protein molecule activation in the semen of infertile patients has been suggested to induce a negative impact on seminal fluid analysis. This study aimed to evaluate Human Peptidoglycan Recognition Protein 1 for indication of bad seminal fluid analysis parameters. One hundred seventy-six infertile patients were included in this study. Seminal fluid analysis and Endtz test were done; Human Peptidoglycan Recognition Protein 1 in seminal plasma using enzyme-linked immune-sorbent assay technology PGLYRP‐1 is evident in seminal plasma of infertile men and statistically different according to the type of infertility. The concentration of PGLYRP-1 in the semen is directly proportional to the presence of leukocytes (Endtz test), which is reflected negatively on the quality of the semen. To the best of our knowledge, this study has not been done in Iraq or elsewhere. The presence of PGLYRP-1 protein in the semen may be evidence of leukocytospermia that may be related to bad quality for semen across infertile men with higher sensitivity than the Endtz test.
Background: Sperm cells are well equipped with a powerful defense system of antioxidants, but an imbalance between the production of reactive oxygen species (ROS) and the available antioxidant-defenses result in oxidative stress. Therefore, antioxidants are supplemented extracellularly under in vitro conditions. Antioxidants are the main defense factors against oxidative stress induced by free radicals. Vitamin E is believed to be the primary component of the antioxidant system of the spermatozoa and is one of the major membrane protectants against ROS and lipid peroxidation (LPO) attack. It appears to be the first line of defense against the peroxidation of polyunsaturated fatty acids (PUFAs) contained in the cellular and sub-cellular membrane phospholipids because of its lipid solubility. Objective: The aim of the current study to evaluate effects of supplementing semen extender with vitamin E, at various concentrations (2, 6 and 12 IU/ml) on sperm motility, survival and morphology on the releases of free radicals and antioxidant enzymes within semen. Patients ,Materials and Methods: Twenty two patients were involved in the present study semen samples were obtained, and sperm count was assessed. The antioxidant (vitamin E) was formulated to be tested at three different levels as follow; 2, 6 and 12 IU/ml; Therefore, control and 3 antioxidant-containing extenders were prepared for semen dilution. The sample was divided into 4 aliquots. One volume of semen was added to 5 equal volumes of the designed extender. Extended semen samples (37˚C) were gradually cooled to 4˚C in the refrigerator and stored for 48 hours. A semen sample was taken out after 48h of storage, then warmed to 37˚C and checked for progressive motility, viability and abnormality. Sperm viability was assessed by Eosin Y. (0.5%)—Nigrosin (0.1%) staining mixture. A total of 200 sperm were assessed under oil immersion with a high-resolution (X100) objective. Sperm morphology was assessed by Hematoxylin-Eosin satin, at least 200 sperm were scored on randomly chosen field, under oil immersion with a high-resolution (X100) objective. Results: The highest post-thaw motility and sperm survival (44% and 51.7%) 48 h after storage at 4˚C (Table 1) were attained (P < 0.01) in the basal diluents containing 6 IU vitamin E. Likewise, the correspondent percentages of live spermatozoa were 59%. Contrariwise, percentage of sperm abnormalities for the previous treatment was 25% significantly (P < 0.05) decrease compares with basal diluents containing 2, 12 IU vitamin E. The lowest (P ≥ 0.05) survival (23.5%, Table 1) was found in the basal diluents containing 12 IU vitamin. Beyond 6 IU vitamin E, there appears to exert adverse effects on sperm viability and survival. Conclusions: For the best protection against the increased free radicals production during chilling-preservation of human semen, extenders must contain 6 IU vitamin E per ml. The intensive production of free radicals in seminal plasma was counteracted by the inclusion of vitamin E at the above mentioned concentrations.