Successful implantation needs a receptive endometrium, a functional and normal embryo at the stage of blastocyst developmental and a synchronized dialogue between embryonic tissues and maternal tissues. The current study was planned and conducted to investigate the potential role of vaginal pessary of hyaluronic acid on intracytoplasmic sperm injection (ICSI) outcome. This study included 63 infertile women (32 women as study group receiving hyaluronic acid and 31 control group) who were subjected to ovulation induction programs with gonadotropin releasing hormone (GnRH) antagonist protocols for in vitro fertilization (IVF)/ intracytoplasmic sperm injection (ICSI) procedures. Blood samples were taken by Vacutainer tubes and centrifuged according to the manufacturer’s instructions for the preparation of serum samples. Serum samples for Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1) were obtained from each patient when the basal hormonal status was determined. The second time point is when the samples were obtained on the day of ova pickup, after that the G1 embryos will be transferred inside the uterine cavity. Then performance of endometrial receptivity parameters or patterns evaluation and patients were followed up to do pregnancy test after14 days after embryo transfer. On the day of human chorionic gonadotropin (hCG), there was no significant difference in mean serum Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1) between hyaluronic acid group and reference group, respectively (p = 0.824). There was no significant difference in mean follicular fluid Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1) between the hyaluronic acid group and reference group, 0.92 ±0.36 pg/ml versus 0.82 ±0.22 pg/ml, respectively (p = 0.223). Treatment with the hyaluronic acid resulted in non-significant increase in serum and follicular fluid Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1), and this increment is not related to Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1) improvement in local blood endometrial blood flow as reflected by reduction in resistive index and pulsatility index with eventual improvement in pregnancy outcome.
Phosphatidylserine (PS), exposed on viable sperm, is required for fertilization as it is recognized by specific receptors located on the microvilli of the oocyte to promote sperm: egg fusion. The heads of viable and motile sperm exhibit PS. With (PS) exposure progressively increasing during sperm transit through the epididymis. PS asymmetry on the leaflets of plasma membrane is arranged in a complex way, via multiple proteins functioning in opposite directions; further, PS is also essential for many intracellular trafficking events. The aim of this study was to evaluate the phosphatidylserine PS level in seminal plasma of infertile men before and after activation using Direct Swim Up (DSU) and Indirect Swim Up (IDSU) activation method and make a comparison between them. Forty infertile men were included in this study during their attendance to the infertility clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University. Normozoospermic or mild asthenozoospermic semen samples were used to choose the cases, according to WHO manual standards (1999 -2010). The collected semen samples obtained were divided into three parts: The first part prepared was as an in vitro sperm characterization before activation while the second and third parts were after activation using direct and indirect swim up techniques respectively. The level of seminal plasma phosphatidylserine was increased after the activation techniques from the baseline level before activation. Comparison of Seminal plasma phosphatidylserine levels between direct and indirect sperm activation technique were significantly higher in direct swim up activation technique than indirect one. Phosphatidylserine (PS) in semen plasma of infertile men was significantly increased after activation using Direct Swim Up DSU preparation method.
Background: Immunological infertility is expected to be the reason of infer-tility in 9-36% of the concerned twosomes. The major cause of immunological infertility is the creation of anti-sperm antibodies (ASA), which influence the capability of insemination of spermatozoa. It has been detected that anti-sperm antibodies (ASA) are present either systemically in blood or locally in seminal plasma of approximately 10% of infertile male patients. Immunity to sperm can cause infertility; humoral antibodies directed against sperm did not necessarily impair fertility unless the circulating antibodies are also present within the re-productive tract and on the living sperm surface. Hormonal imbalance and sperm autoimmunity have been considered as two systems that play in near association and affect each other. Testosterone is the Steroid hormone necessary for the de-velopment and maintenance of secondary sexual characteristics as well as initi-ation and maintenance of Spermatogenesis .It was known that males with abnor-mal seminal fluids have lower concentrations of the testosterone hormone. Objectives: To study the serum & seminal plasma antisperm antibodies levels in relevance to levels of testosterone in serum and seminal plasma accordingly to sperm function parameters in different groups of infertile patients. Subjects,Materials and Methods: Blood and semen samples were collected from (80) subjects (60) infertile patients (20Asthenozoospermic, 20 Oligozoospermic,20Azoospermic) and 20 Normozoospermic subjects .The anti-sperm antibody and testosterone levels were measured by using Enzyme Linked Immunosorbent Assay (ELISA). Results: The Antisperm antibodies were positive in serum of normozoospermic, asthenozo-ospermic, oligozoospermic and azoospermic men respectively are about 10%, 5%, 20%,5% and in the total group was 10% positive. The ASA in seminal plasma of normozoospermic men, asthenozoospermic men, oligozoospermic men and azoospermic men respectively are 0%, 0%, 5%, 5% and in the total groups 2.5%.These findings lead to no possibility to make clear correlations with the studied hormones. There was significant increase (P<0.05) in the levels of testosterone in serum and seminal plasma in comparison between normozoospermic and infertile men subgroups. There was significant decrease (P<0.05) in the levels of pro-gesterone in serum and seminal plasma in comparison between normozoospermic men and infertile men subgroups, the highest levels were observed in asthenozoospermic subgroup. The study showed that 10 patients (12.5%) out of 80 patients showed a positive ASA positive in serum or seminal plasma of all groups. The result was considered positive if the value was ≥60 RU / ml .On these basis and regarding that screening criteria the positive ASA group showed the association with the low levels of testosterone in both types of the samples(serum and seminal plasma), these results was exhibited a high significant difference when compared with negative ASA group. Conclusion: Determination of the relevance of the levels of testosterone and anti-sperm anti-bodies in the serum and seminal plasma, which in turn is important to determine the type of infertility especially the immunological type.
Oxidative stress in the field of reproduction has been extensively considered in association with sperm properties and male infertility; in any case, the effect of oxidative stress in association with oocyte trademark got less thought than that of sperms. The goal of the study is to understand the possible association among serum and follicular oxidative markers in infertile women and oocyte characteristics and pregnancy result in ICSI cycles. The present case-control study included 57 infertile women undergoing ICSI that was done in the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies at Al-Nahrain University. The assessment started in April 2018 and connected with March 2019. Mean metaphase II oocyte count of all women was 5.70±3.87, that of pregnant women was 6.90±3.68 and that of non-pregnant women was 4.46±3.73; there was a basic qualification in mean metaphase II oocyte check among pregnant and non-pregnant women (P = 0.016); being higher in pregnant women. The mean abnormal oocyte count of all women was 0.63±1.26, that of pregnant women was 0.28±0.45 and that of non-pregnant women was 1.00±1.68. Oxidative stress markers inside and out affected clinical pregnancy result (P < 0.05) and appear to impact clinical pregnancy bring about women undergoing ICSI due to its effect of oocyte characteristics.
Pro-fusion proteins as mitofusins-1 are required for controlling mitochondrial shape which determined by a dynamic balance between organelle fusion and fission and also supports oocyte development. When compare with somatic cell, mitochondria of oocyte are tiny and circular in presence. The aim is to study the mitofusin-1 in the follicular fluid as a marker for evaluating of oocyte maturation in women undergoing ICSI cycles. Fifty infertile females were included in cross-section study was undergoing ICSI procedure with age 20 to 42 years. After retrieval of oocyte and the number of oocytes was recorded by embryologist and follicular fluid Samples were used for the measurement of Mitofusin 1. Mitofusin 1 levels by ELISA kit (Mybiosource /USA).The results showed considerable higher mitofusin-1levels in patients with good oocytes quality (3.71 ± 1.35 vs. 2.45 ± 1.12 & p=0.001). There were much positive correlations between follicular fluids mitofusin-1 with both total oocytes count (r= 0.374 & p= 0.007) and MII oocytes (r=0.383 & p=0.006); and there was no much correlation between mitofusin-1 levels with MI oocyte (r= - 0.100 & p= 0.490), germinal vesical oocyte (r=0.103 & p= 0.475) and fertilization rate (r= 0.054 & p= 0.711). High follicular fluid levels of Mitofusin 1 may positively impact oocyte development and pregnancy rate.
Background: The procedures of assisted reproductive technologies have been developed during recent years and have been revealed the need for new suitable and effective techniques of sperm treatment in the laboratory. Objectives: This study is designed to normozoospermic and asthenozoospermic compare the semen outcomes of two in vitro sperm activation techniques and evaluate their efficiency, namely; Glass wool filtration technique and a new technique of Glass wool filtration combined with Pentoxifylline. Subjects, Materials and Methods: Fifty five semen samples were collected from the patients, fifteen were obtained from normozoospermic men, forty were obtained from asthenozoospermic men who at tend the infertility clinic were involved in this study. Sperm parameters were assessed according to WHO (2010 and 1999). Post-activation of each sample were divided into two aliquots, the first one : using Glass wool filtration technique , the second : using Glass wool filtration technique with Pentoxifylline. Results: A significant increase of sperm motility, progressive sperm motility and morphologically normal sperm when using Glass wool filtration technique combined with Pentoxifylline as compared to Glass wool filtration technique. Regarding sperm agglutination and Round cells count the study showed the best results by decreasing their levels when using Glass wool filtration technique combined with Pentoxifylline as compared to Glass wool filtration technique alone. Conclusions: From the results of the present study, the sperm function parameters outcome when using glass wool filtration technique combined with Pentoxifylline were superior to other technique since it yields better sperm function parameters rather than the outcome of glass wool filtration ; respectively especially when using a low quality of semen samples such as decreased sperm function parameters (sperm motility) so it was fit in cases of asthenozoospermia which taken as an example of infertile group within this study.
The study aim is to evaluate the predictive rate of positive sperm retrieval of serum concentration of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (TT) in azoospermic patients presenting to High Institute of infertility diagnosis and assisted reproductive technologies. The study records of 65 azoospermic men with who underwent open testicular sperm extraction between February 2017 to April 2019 were studied. The patient’s age, hormonal profile for FSH, LH and TT and testicular histopathology were recorded. The sperm retrieval rate according to histological findings norm-spermatogenesis (NS), hypo-spermatogenesis (HS), maturation arrest (MA) and Sertoli cell-only syndrome (SCOS) were recorded as follows: 100%, 75%, 44.4%, and 28.6% respectively. The cutoff value of FSH was (≤ 14.56 mIU/L) with an accuracy level of 73.0%, a sensitivity (66.7 %) and a specificity (79.3 %). The cutoff value of the serum level of LH was (≤ 9.37 mIU/L) with an accuracy level of 78.4%, a sensitivity level of 77.7 % and a specificity level of 72.4 %.
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.
There is a relation between adipose tissue and macrophages. These macrophage cells influence adipose tissue by releasing many bioactive chemicals, including the wingless-type MMTV integration site family 5a (wnt5a) glycopeptides and the more traditional cytokines. Healthy adipocytes can release the wnt5a inhibitor Secreted frizzled-related protein 5(SFRP5). However, this protective effect was found to be reduced in obesity. This study aimed to assess the effect of follicular and serum levels of SFRP5 as well as serum and follicular levels of wnt5a on ICSI outcome and determine the potential roles of SFRP5 as a predictor of positive pregnancy in Iraqi infertile females undergoing ICSI with different body mass index .Ninety infertile females who received intracytoplasmic sperm injection and had various infertility factors between 18 and 45 years participated in this prospective clinical cross-sectional study. Follicular fluid, serum SFRP5, and serum and follicular fluid wnt5a levels were measured for all participants on the day of ovum pickup. In addition, ICSI outcomes were determined. There was a significant difference in the mean total gonadotrophin dose used among the three studied groups (p = 0.023), higher in overweight and obese females than normal-weight females. There was a positive significant correlation between serum plus follicular fluid wnt5a and total gonadotropin dose. Serum and follicular fluid SFRP5 levels were significantly higher in normal-weight females. There were higher serum (p-value=0.002) and follicular fluids (p-value=0.005) SFRP5 levels in pregnant females. Both SFRP5 and wnt5a glycoproteins were detected in infertile women's serum and follicular fluid. Serum and follicular fluid secreted frizzled-related protein 5 level positively associated with ICSI outcome. The cutoff values for serum and follicular fluid SFRP5 were ≥15.86 (ng/ml) and ≥ 11.3(ng/ml), respectively, as predictors of positive pregnancy in women undergoing IVF/ICSI cycles. Obesity, BMI, SFRP5, wnt5a, ICSI Outcome
Hyaluronic acid (HA) is a polysaccharide, that composed in the extracellular matrix of the cumulus cells, and considered to take function in choosing mature spermatozoa. The Sperm-hyaluronic binding assay (HBA) is significant diagnostic instrument for suspected male infertility when examining the semen. The HBA slide supplies the ratio of mature binding sperm in the specimen. To evaluate sperm parameters, and to evaluate the (HBA Score percentage (%) before and after activation. Twenty-five infertile couples a sharer in this current study through their attendance to the Al Nahrain University. Males undergoing seminal fluid analysis were done according to (WHO 2010, and WHO 1999). Sperm parameters assessed and calculate the HBA Score% pre-and-post-direct swim-up technique, the results were statistically tested. In Post-activation there was considerable increment noticed in the progressive motility and morphology of spermatozoa, likewise significant improvement in the (HBA score %). Furthermore, considerable reduction in the concentration, agglutination of spermatozoa and round cells count, when compared to pre-activation. The mean of binding of spermatozoa to hyaluronan pre-activation (direct swim-up technique) was less than normal limit for normozoospermic males. However, it significantly improves post-activation.
Obesity induces metabolic dysfunctions in bodily systems and is a significant health challenge. Excessive adipose tissue accumulation encompasses dynamic, endocrine, and biologically active processes. The fundamental pathogenic mechanism of obesity is chronic low-grade inflammation exacerbated by pro-inflammatory biomarkers such as free fatty acids, lipopolysaccharides, adipokines, and cytokines. The rising prevalence of obesity over the past forty years has led to an increase in associated comorbidities, including polycystic ovarian syndrome (PCOS). To clarify the impact of increasing body mass index (BMI) in PCOS and non PCOS on the ICSI outcomes of 143 infertile Iraqi females, dividing them into two main groups: PCOS and non PCOS and each group into normal weight and overweight/obese group. Group 1= normal BMI PCOS, group 2= overweight/ obese PCOS, group 3= normal BMI non PCOS, group 4= overweight/obese non PCOS. Significant differences in ICSI outcomes were observed in the PCOS groups compared to the non-PCOS groups concerning the total number of harvested, MI, MII oocytes, GV, abnormal and fertilised oocytes, total and D3 grade I embryos alongside higher number of arrested and transplanted embryos. There are numerically higher number of D3 grade II and III embryos, morula and blastocyst with no significant difference seen. The rates of maturation and fertilisation exhibited no significant disparities. The biochemical pregnancy rate was higher in group 2 compared to group 1, and in group 4 compared to group 3, with a statistically significant difference. A higher BMI among younger PCOS individuals may be associated with better pregnancy outcomes.
Plasminogen Activator Inhibitor-1 expression in serum and follicular fluid plays a crucial role in regulating fibrinolysis during the pregnancy period by blocking the action of other proteins called plasminogen activators. The aim is to assess the Plasminogen Activator Inhibitor-1 level in blood serum and follicular fluid as a predictive marker of pregnancy. This study included Sixty-nine infertile females undergone ovarian stimulation with antagonist protocol and received recombinant follicle-stimulating hormone, at the cycle day two. The serum and follicular fluid were collected from each female on the day of oocyte retrieval, for PAI estimation using the Elisa kit. The data were analyzed using version 23.0 SPSS and Microsoft Office 2010. The serum PAI-1 levels at the day of oocyte retrievals were significantly lower in positive pregnancy women in comparison with non-pregnant women (p < 0.05), Yet no significant follicular fluid PAI-1 levels were noted in comparison with non-pregnant women. There was a negative significant correlation (p > 0.05) between serum and follicular fluid PAI-1 levels on the day of oocyte retrieval according to oocyte ICSI outcome. The evaluation of serum PAI-1 levels can be used as a promising parameter for the prediction of pregnancy outcomes of ICSI patients. It is assumed that the pregnancy rate was affected by PAI-1 level during ICSI cycles. The concentration of PAI-1 in serum and follicular fluid at the retrieval day is not correlated with the oocyte's maturity.
Infertility has been defined as the disability to fulfill pregnancy after twelve months of regular sex coition, with no contraceptive taken. Male factor infertility was explained in terms of low spermatozoa concentration (oligozoospermia), or morphology (teratozoospermia), impaired motility (asthenozoospermia), or in some cases, total lack of spermatozoa in the ejaculate (azoospermia). In several cases, a collection of one, or more of these sperm variables defects may be noticed, evaluation these factors by the initial and most important step is seminal analysis. Our study aims to study some sperm characteristics in asthenozoospermia men in comparison with normozoospermia men before and after glass wool and Sephadex activation. This study involved 60 semen samples collected from male patients that came to male infertility clinic at Al-Nahrain University; the recruited semen samples were divided into 2 groups, (40 asthenozoospermic and 20 normozoospermic subjects). After collected semen samples, and assessed analysis of seminal fluid. Each semen samples were divided into 3 aliquots. The first aliquot prepared was In-Vitro to assessed sperm characterization before activation, the second part using glass wool filtration (GWF) technique, while the last part was prepared using Sephadex. Both techniques resulted equally in reducing sperm concentration, were equally effective in upgrading sperm motility and in minimizing round cell count; however, Sephadex was superior to glass wool method in upgrading the percentage of morphologically normal sperm. Both Sephadex and glass wool techniques has been proved effective to improve semen quality.
Due to malfunction of the hypothalamus-pituitary-ovary axis and probable dysregulation of ovarian activities, ovary has a large number of small follicles but no developed follicles. L-Carnitine is a tiny, water-soluble chemical that plays a critical role in fat metabolism. It is required for appropriate mitochondrial fatty acid oxidation and acyl-CoA ester excretion, as well as affecting ATP levels. The current study was aiming at evaluating the protective effect of L-Carnitine (LC) against deleterious substances present in serum of patients with PCOS, which may affect the oocyte maturation. 60 infertile women diagnosed with polycystic ovarian syndrome have been enrolled in this study and enter their Intracytoplasmic Sperm Injection (ICSI) cycle; they were informed about the study and signed a written informed consent. They were grouped into treatment group (n=30) who were treated with L-Carnitine® tablet for two months and placebo group (n=30). All enrolled women underwent flexible antagonist protocol for ICSI. Significant increase in mean mature metaphase II (MII) oocyte count in study group than placebo group, oocyte maturity rate was significantly higher in study group in comparison with placebo group. In addition, mean fertilized (2PN) oocyte count was increased significantly in study group in comparison with placebo group. Mean fertilization rate was significantly higher in study group in comparison with placebo group. Treatment with of patients with polycystic ovary syndrome of L-Carnitine resulted better oocyte quality and the improvement in oocyte quality is reflected by mature MII oocytes, maturity rate and fertilized rate with eventual improvement in embryo quality.
Polycystic ovarian syndrome (PCOS) is a group of clinically distinct endocrine conditions. Insulin resistance-related proteins are involved in PCOS pathogenesis. Irisin is a new myokine that functions similarly to adipokines. Irisin has been linked to insulin resistance and metabolic syndrome. There have been conflicting findings regarding irisin levels and their connection to hormonal and metabolic alterations in PCOS .This study aimed to compare the levels of irisin in follicular fluid and serum between women with PCOS and healthy women and correlate them with oocyte and embryo quality in women undergoing the ICSI cycle .Ninety infertile Iraqi women were enrolled, 46 women with PCOS and 44 weight-matched non-PCOS controls, in whom ICSI was done for them and evaluated the level of follicular fluid, serum Irisin, and insulin level with measure of Homeostasis Model of Assessment-Insulin Resistance (HOMA-IR) and correlate them to with oocyte and embryo quality. HOMA-IR, FF, and serum irisin levels in PCOS considerably increased. (P ≤ 0.01). In addition, there was a significant negative correlation between Irisin levels in follicular fluid and serum with MII oocyte, oocyte maturity rate, and grade 1 embryo. The serum, follicular fluid Irisin, and HOMA-IR were significantly higher in PCOS than in control cases, which may cause a reduction in oocyte maturation, embryo quality, and pregnancy outcome.
Background: One of the major concerns among the male reproductive health is the inability to achieve parenthood even after regular unprotected intercourse. The efficiency of the sperm selection techniques is expressed as the concen-tration of spermatozoa with normal motility. Discontinuous density gradient centrifugation (DGC) method widely used over the last decade due to effec-tiveness and reproducibility in recovering most motile sperm, which separates sperm based on their motility, size, and density differential. Glutathione act as an amino acid donor during spermatogenesis. If the levels of glutathione are too low during spermatogenesis, the number of mature and morphologically normal spermatozoa generated will be decreased. Objectives:The aim of this study was to assess the beneficial role of TAD 600mg Glutathione combined with DGC technique for activation of human sperm of asthenozoospermic, oligozoospermic and normozoospermic subjects compared with the DGC technique alone. Patients, Materials and Methods:Sixty males were involved in this study, di-vided into three groups, (twenty asthenozoospermic, Twenty oligozoospermic and twenty normozoospermic subjects)during their attendance to the Infertility Clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Tech-nologies; Al-Nahrain University. Semen samples were obtained, and seminal fluid analysis was assessed. Each semen sample was divided into three parts. The first part prepared as in vitro sperm characterization before activation, the second part using density gradient centrifugation technique, while the last part prepared using density gradient centrifugation combined with TAD 600mg Glu-tathione. Results: After in vitro sperm activation for asthenozoospermic, oligozoospermic and normozoospermic samples a significant increase was observed in the sperm function parameters including sperm concentration, motility and morphological-ly normal sperm percentage as compared to pre-activation using combined tech-niques (density gradient centrifugation and TAD 600mg Glutathione) as com-pared to DGC technique alone. Conclusions:TAD 600mg Glutathione combined to the DGC technique was found to give higher significant results on sperm function parameters (sperm concen-tration, motility and morphology) when using a low quality of semen samples such as decreased sperm motility as compared with DGC technique alone.