Background: Diabetes has been found to induce various subtle molecular changes important for sperm quality and function: The mammalian spermatozoa are rich in polyunsaturated fatty acids, making them particularly sensitive to the deleterious effects of lipid peroxidation, which may result in irreversible loss of motility and a low level of fertilizing capacity. An antioxidant, glutathione (GSH) a polypeptide is important in biological oxidation-reduction reactions and improves sperm motility. Objective: The study was aimed to investigate the effects of GSH injection on healthy and diabetic male mice fertilization potential in vitro following in vitro fertilization (IVF). Materials and Methods: Inducing diabetes in mice through intra peritoneal alloxan injection. The treatment with GSH divided into two periods 10 and 20 days, studying the in vitro fertilization outcomes. Results: Glutathione daily IV injected in a period of 10 to 20 days for the diabetic induced male mice significantly increase the sperm concentration and motility and decrease the abnormal and dead sperms percentages in treated group compared to untreated group. The Fertilization rate was significantly (P<0.05) increased in mice treated with GSH compared to untreated one. Conclusion: Glutathione injections may decrease the oxidative stress caused by diabetes and enhance the in vitro fertilization potentials in male mice.
Polycystic ovary syndrome (PCOS) is a common cause of anovulatory infertility and is associated with an increased risk of premature luteinizing hormone (LH) surges during controlled ovarian stimulation. Progestin-primed ovarian stimulation (PPOS) using Medroxyprogesterone Acetate (MPA) has emerged as a convenient alternative to conventional gonadotropin-releasing hormone (GnRH) antagonist protocols. This study aimed to compare the effects of MPA-based PPOS and GnRH antagonist protocols on oocyte yield, maturation, fertilization, and embryo quality in women with PCOS undergoing ICSI . This prospective, randomized, open-label controlled clinical trial included 108 women with PCOS undergoing ICSI at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, Baghdad, Iraq, between 2023 and 2024. Participants were allocated to an MPA-based PPOS group (n=49) or a flexible GnRH antagonist group (n=59). Oocyte yield, metaphase II (MII) oocytes, fertilization rate, embryo quality, ovarian sensitivity index (OSI), gonadotropin consumption, stimulation duration, and LH profiles were evaluated.No significant differences were observed in baseline characteristics or hormonal profiles between groups. The MPA group showed a numerically higher number of retrieved oocytes (20.65 ± 7.50 vs. 13.39 ± 4.41; p=0.083) and MII oocytes (15.10 ± 5.20 vs. 11.24 ± 4.12; p=0.09), although these differences were not statistically significant. Fertilization rates were comparable between groups (72.4 ± 10.6% vs. 71.2 ± 11.3%; p=0.68), as were embryo quality, OSI, gonadotropin consumption, and stimulation duration. Both protocols effectively suppressed premature LH surges without premature ovulation or cycle cancellation. MPA-based PPOS appears to be an effective and practical alternative to GnRH antagonist protocols for PCOS patients undergoing ICSI, providing comparable fertilization and embryological outcomes with effective LH suppression. Its oral administration and potential cost advantages may be particularly valuable in resource- limited ART settings.
Background: Fibrillin- 3 is a recently discovered hormone.However, there are no studies till now regarding the role of fibrillin-3 hormone on fertilization capacity in infertile pregnant and non-pregnant women. Objectives: The goals of this study are concentrated on the fertilization capacity parameters by examining the effect of in vivo concentration of fibrillin-3 hormone on the ovarians status at different menstrual cycle phases and to elucidate the correlation between the levels of this hormone with pregnant and non -pregnant women following Intrauterine insemination (IUI) outcome or through natural pregnancy. Methods: Sixty infertile couples were involved in this prospective study. The couples were divided according to the type of insemination and ovulation induction program into two groups Group(1): Thirty spouses were subjected to induced ovulation stimulation by using clomiphene citrate(CC) and recommended for natural coitus. Group(2): Thirty couples, the females were induced ovulation stimulation by CC, Gonal F-751U /ml and Fostimon 50IU/ml with IUI. Measurements of Fibrillin-3 with other reproductive hormones were done at different phases of menstrual cycle and for pregnant and non-pregnant women following natural coitus and IUI. Results: The mean of fibrillin-3 hormone level at day 2 of menstrual cycle was 3.7ng/ml (ranged from 3.50 to3.91) in the two groups. The mean of fibrillin-3 hormone at cycle day 13 was 6.6ng/ml( ranged from 5.83to 7.0). Whereas, The basal level of Fibrillin-3 at 14 days post insemination was 3.3ng/ml. The range is from 2.76,395 to 3.29 ng/ml. There was no significant difference in the mean of Fibrillin -3hormone between pregnant and non-pregnant women through the phases of menstrual cycle or between the two groups, but there is a significant value between fibrillin-3 post insemination and E2 day 2 and day 13 P<0.05. Conclusion: The present study concluded that Fibrillin -3 hormone has no role in pregnancy, but it may have a main relationship with estrogens reaction in folliculogenesis, oocyte maturation and follicular contents. Therefore, this hormone may be one of the new hormones that interfere with fertilization capacity.
Background: In vitro Fertilization (IVF) is an important alternative for infertile couples, there is an increase in the number of children conceived by IVF worldwide. Objective: The aim of this study was to assess the risk of birth defects and adverse outcome among infants and children conceived by in vitro fertilization in Sulaimani-Iraq. Patients and Methods: This case-control study was carried out on 160 children, 75 were product of IVF(using ICSI) and 85 were product of natural conception. Data collected regarding mother age, father age, duration of infertility before pregnancy with the index child, any complication during pregnancy, age of the child, gender, mode of delivery, gestational age, weight at delivery, history of neonatal admission and it’s cause, admission to hospital after neonatal period, and history of surgical operation. All children were examined for any gross anomaly, cardiovascular, respiratory, gastrointestinal, skeletal, urological and neurologic abnormality. Results: IVF was a risk factor for prematurity (OR, 7.844 95% CI 3.025 - 20.339) , LBW (OR, 4.645 95% CI 2.038 -10.587), Congenital anomaly (OR, 1.362 95% CI 0.720 - 2.576), predominance of male gender (OR, 1.340 95% CI 0.719-2.498), neuro-developmental delay(OR,1.333 95% CI 0.672-2.646), admission to NICU (OR,1.703 95% CI 1.052-2.758 ), C/S delivery (OR, 1.972 95% CI 1.507-2.581), and childhood hospitalization (OR, 1.193 95% CI 0.838-1.696), also there was statistically significant association with multiple pregnancy. Conclusions: IVF carries increased risk of LBW, prematurity, congenital anomaly, neuro-developmental delay, admission to NICU, childhood hospitalization, C/S as a mode of delivery and predominance of male gender.
Activin is a member of the transforming growth factor-beta superfamily and is a polypeptide growth factor. Activin regulates follicle growth, gonadotropin responsiveness, steroidogenesis, oocyte maturation, ovulation, and corpus luteum function in the ovary via a local autocrine/paracrine effect. Activin measurement A level in serum and follicular fluid during the preovulatory stage in PCOS women undergoing an IVF/ICSI cycle to predict fertilization capacity earlier Fifty infertile couples, 25 with PCOS and 25 without PCOS, who attended Al Nahrain University in Baghdad, Iraq's High Institute of Infertility Diagnosis and Assisted Reproductive Technologies. The females that participated were between the ages of 18 and 40. On the day of oocyte retrieval, the level of Activin A in serum and follicular fluid was measured. Activin A levels in PCOS women's serum (activin A=1274 77.9) and follicular fluid (activin A =1038 50.3) compared to non-PCOS women's serum (activin A=453 25.74) and follicular fluid (Activin A=450 23.73). The study shows no correlation between activin A hormone with oocytes characteristics and fertilization rate in PCOS women with a significant increase of activin A hormone in the serum and FF of PCOS women compared to non PCOS women. Activin A level can predict a diagnosis of PCOS in addition to positive predictive pregnancy but not indicator of fertilization capacity in PCOS women.
Background: Zygote produce from once a sperm fertilizes an egg cell. Then, the zygote (unicellular) will begin chain of cellular cleavages to produce multicellular mass, its embryo, the differentiated to different tissues and organism. The development of the embryo is called embryogenesis. Coenzyme Q10, is an antioxidant produced in the body. It boosts cellular energy and may enhance the immune system. CoQ10 is present and measurable in seminal fluid, the concentration of CoQ10 directly correlates with both sperm count and motility. It is beneficial in the prevention and treatment a wide range of health problems. Objectives: The present study was aimed to investigate the possibility of using coenzyme Q10 to improvein vitro fertilization (IVF), and early embryonic development (ED) in mice as a model for human being. Methods: Superovulation program was achieved to mature healthy female mice with age 10-12 weeks and weight 24-26 gm. After sacrificing female, oocytes were collected and incubated within CO2 incubator for less than 1 hour. Sperm were collected from vas deference of males. Sperm parameters were assessed after 30 min. ofincubation. Mature oocytes were divided into three groups according to theconcentrations of CoQ10 including G1 (control group; SMART medium only), G2 (treated group; SMART medium enriched with 20 M CoQ10) and G3 (treated group; SMART medium enriched with 40 M CoQ10). IVF technique was performed for 3 groups, and assessment of IVF (%), embryonic development stage (%) and abnormal embryo morphology (%) for each embryo stage. Results: Results of the present study appeared significant increment (P<0.05) in the percentages of IVF for both treated groups as compared to the control groups. Also, significant increase (P<0.0P) in the IVF (%) was observed when using 40 M CoQ10 as compared to 20 M CoQ10. Non significant differences (P>0.05%) in the 8-cells embryo stage were assessed among control and treated groups. Conclusion: From the results of the present study it wasconcluded that the coenzyme Q10 (40µM) enriched to the culture medium improved percentage of in vitro fertilization and no effect on embryonic development.
Background: Citrullus colocynthis(CC) is an herbal medicine used in the treatment of a wide range of diseases. Recently, the effect of this plant on the reproductive system has been studied.However, its role in vitro still unclear. Objective: The present study was designed to investigate the possibility of using CC extract for in vitro sperm direct activation technique, in vitro fertilization(IVF) and early embryonic development using the mice as a model for mammals. Materials and Methods: Citrullus colocynthis extract (0.05mg/ml culture medium) was used for in vitro direct sperm activation technique. The same medium was used for oocytes insemination and for culturing the embryos after 24-48 hours of insemination. The oocytes were collected from superovulated female mice and divided into two groups: first group, 343 oocytes were inseminated and cultured in CC- free Ham's F-12 medium (the control group). The second group, 345 oocytes were inseminated and cultured in 10% CC - Ham's F-12 medium (treated group). Each 4 oocytes were inseminated with the same sperm concentration (1-2 x 10^5 sperm/IVF well). The fertilization rate was recorded after 24 hours of insemination, while embryonic development rate was recorded after 24 and 48 hours of insemination. Results: In vitro activation of epididymal sperms with 10%CC has shown positive effect on sperm concentration, sperm motility, and grade activity of progressive forward movement. There was a highly significant (P<0.004) increase in FR of the treated group (61.5%) compared to control group (50.4%) after 24 hours of insemination. Embryonic developmental rate was significantly increased after 24 and 48 hours of insemination in treated group compared to control group. Conclusion: It concluded that adding the 10%CC to the culture medium of the epididymal sperm and in vitro inseminated lead to improvements in certain sperm function parameters and sustain the FR and early embryonic development rate.
Back ground: Optimal IVF and embryo culture conditions depend on the composition of culture media being the most important determinants of successful in vitro interaction of the gametes and subsequent embryo development. ATP is fundamental factor to maintain the life, by providing energy, and controlling the cell function and metabolism. Objective: To investigate the effect ATP supplied to culture medium on fertilization rate and in mice as model for human being. Materials and Methods: The female mice (90) were superovulated using superovulation program (SOP) to produce large number of oocyte were divided into three groups, G1 inseminated with vassal sperm in vitro alone (control) , G2 inseminated with vassal sperm in vitro using SMART enriched with ATP(1.25mM). While, G3 inseminated with vassal sperm in vitro using SMART medium enriched with ATP (2.5mM). Mouse spermatozoa were collected from both vas deferens. Then sperm parameters were assessed after 30 min. IVF technique was performed for 3 groups. Results: The results of the present study appear, significantly increment (P<0.05) in the IVF (%) was observed when using 2.5 mM ATP as compared to 1.25 mM ATP and control group. Conclusions: The result showed that addition of high ATP concentration (2.5 mM) enriched to the culture medium improved percentage of in vitro fertilization.
Background:Defects in reproduction that may involve both the male & female partners result in the inability to procreate which is known as infertility. The use of assisted reproductive technologies (ART’s) has been increasing over the past three decades. Many efforts have been done to compensate for unsatisfac-tory results in in-vitro fertilization (IVF) procedures, so that patients undergo different ovarian stimulation protocols to control retrieval of multiple oocytes in a single cycle. Although ovarian stimulation has an important role in ART’s, it may also have detrimental effects on oocyte quality, embryo quality, and IVF outcomes. Objectives: To assess the effect of long agonist versus antagonist protocols on oocyte and embryo quality in IVF-Intracytoplasmic sperm injection (ICSI). Patients,Materials and Methods: This comparative study was done on the embryos of seventy one infertile fe-males who were undergoing Intracytoplasmic sperm injection (ICSI) treatment regardless to the presence or absence of previous trials. These patients were divided into two groups, 45 of them treated with agonist protocol and other 26 patients treated with antagonist protocol. First oocyte retrieved was used for further assessment. Of the 71 oocytes, there were eight immature oocytes (3 at germinal vesicle (GV) stage, 5 at metaphase I (MI) stage) and three abnormal oocytes, and these were excluded from the following ICSI treatment. Assessment and comparison of oocyte maturity, fertilization rate, cleavage rate and resulting embryo quality were done. Results:The results of the present study showed significant difference (p<0.05) in oocyte number and maturity but no significant (p>0.05) difference between the group treated with long agonist and those on antagonist protocol regarding fertilization rate, cleavage rate, and embryo quality. Conclusions:The type of protocol used in in-vitro fertilization has no detrimen-tal effect on oocyte or embryo quality.
Background: Many studies found that the Glycyrrhiza glabra (G.glabra) showed a positive effect on the activation of sperm in vitro specifically improve the activity of progressive movement. However, it was not known the impact of this stimulant on the certain sperms parameters in vasal obstructive mammals. Objectives: This experiment was designed to examine the possibility of fertilization ova by epididymal sperm of vasectomized mice activated by G. glabra and study the early embryonic development, as a model for obstructive azoospermia. Methods: Glycyrrhiza glabra aqueous extract (0.3 mg/ml culture medium) was used for in vitro direct sperm activation technique. The superovulated female mice (SUO) were divided into two groups, the first group; the insemination of flushed ova was performed with epididymal sperm of vasectomized male mice activated by adding30% of G. glabra to the culture medium (treated group).While the second group,the ova were inseminated by epididymal sperm of vasectomized male mice activated by G. glabra - free medium (control). Results: This study showed a highly significant (P <0.01) improvement in certain sperm function parameters i.e the sperm forward movement, morphologically normal sperms (MAS). There was a significant (P<0.05) increase in fertilization rate (FR) of SUO mice oocyte using 30% G. glabra medium(75.84%) compared to G. glabraFree medium (Hams -F12 medium) alone (54.43%).By adding 30% G. glabra to the medium, the number of 3-cell and 4-cell embryonic stage of SUO mice was significantly (P <0.05) higher than that of grouping G. glabra- Free medium mice embryo(62.10% and 65.78% vs. 58.88 and 66.29%, respectively). Conclusion: This study showed a highly significant (P <0.01) improvement in certain sperm function parameters i.e the sperm forward movement, morphologically normal sperms (MAS). There was a significant (P<0.05) increase in fertilization rate (FR) of SUO mice oocyte using 30% G. glabra medium(75.84%) compared to G. glabraFree medium (Hams-F12 medium) alone (54.43%).By adding 30% G. glabra to the medium, the number of 3-cell and 4-cell embryonic stage of SUO mice was significantly (P <0.05) higher than that of grouping G. glabra- Free medium mice embryo(62.10% and 65.78% vs. 58.88 and 66.29%, respectively).
To explore whether dual triggering for final oocyte maturation using a low dose of recombinant human chorionic gonadotropin (hCG) plus gonadotropin-releasing hormone agonist (GnRH-a) is as effective as the standard dose of recombinant hCG triggering alone in term of a total number of retrieved oocytes, fertilization and pregnancy rate in fresh autologous antagonist intracytoplasmic sperm injection (ICSI) cycles. 80 infertile women undergoing ICSI, aged ˂ 40 years, 18˂ Body mass index ˂30 Kg/m2 included in fresh autologous antagonist ICSI cycles. 40 women received a Standard dose of recombinant hCG (500 µg) alone and 40 women received GnRH-a (0.2 mg triptorelin plus 250 µg recombinant hCG) were used for final oocytes maturation. This study was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Techniques, Al Nahrain University. The mean total number of retrieved oocytes, fertilization, and pregnancy rate were measured. The mean total oocyte number being statistically higher in the dual trigger group. The mean number of fertilized oocytes and clinical pregnancy rates was higher in the dual trigger group than in the control group, but the difference showed borderline statistical significance. Furthermore, fertilization rates show no statistical difference between the control group and the dual trigger group. Results of the study confirm that dual triggers show favorable improvement in fresh autologous antagonist ICSI cycles with the use of GnRH agonists plus hCG compared with hCG alone for triggering of final oocyte maturation.
Inhibition of luteinizing hormone (LH) by gonadotropin releasing hormone (GnRH) antagonist may lead to suboptimal response during ovarian stimulation. In addition, several studies suggest that low level of LH is associated with lowered fertilization and implantation rate and increased early pregnancy loss rate. The aim of this study is to study the effect of adding low dose human chorionic gonadotropin (hCG - 200 IU), as an LH supplement, to recombinant follicle stimulating hormone (rFSH) in a GnRH antagonist cycles in women undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment. Sixty-three infertile women undergoing IVF were randomly divided into two groups. One group was stimulated with the conventional stimulation protocol (rFSH alone), while the second group received 200 IU hCG in addition to rFSH in the late follicular phase (hCG + rFSH). Both groups' results including pregnancy rate, total dose of rFSH required, duration of stimulation, endometrial thickness, oocytes and embryos characteristic, serum hormone levels (Testosterone, Estradiol, Progesterone, and LH) and level of epidermal growth factor (EGF) were compared. The results show that pregnancy rate among the group of women who received the low dose hCG was higher than those who did not receive hCG. However, this difference did not reach statistical significance. Furthermore, other cycle outcomes and hormonal values were comparable between the two stimulation protocols.
The estimation of oocyte quality in human in vitro fertilization (IVF) is an important point for the embryologists. Oocyte selection and the identification of the best oocytes may help to limit an overproduction of embryos and to improve the results of oocyte cryopreservation programs. The follicular fluid (FF) can be provided easily during oocyte pick-up and known to represents an optimal source for non-invasive biochemical predictors of oocyte quality. However, till now no substance was found to be used as reliable markers of oocyte competence to fertilization, embryo development and pregnancy. Metabolism and ATP levels within the oocyte and adjacent cumulus cells are associated with quality of oocyte and optimal development of a healthy embryo. Lipid metabolism provides a potent source of energy and its importance during oocyte maturation is being increasingly recognised. Objectives: To determine the effect of lipids (Cholesterol. Triglyceride and HDL) level in serum and follicular fluid in predicting oocyte quality, embryo quality and outcome in patients undergoing intracytoplasmic sperm injection program Patients and Methods: Fifty eight (58) infertile women with an age range 21-41 years old undergoing intracytoplasmic sperm injection cycles were included in this study. Lipids (Cholesterol. TG and HDL.,) levels in serum and follicular fluid were measured by auto analyzer for biochemistry (Flexor El_200) and correlation with oocyte quality, embryo quality and ICSI outcome was done. Results: The results of this study showed no significant (p<0.05) difference in lipid levels in serum and follicular fluid between pregnant and non pregnant females as well as non significant correlation was found between lipid level in serum and follicular fluid in relation to oocyte quality and embryo quality. Conclusions: Serum and follicular fluid lipids are not good predictors of oocyte quality, embryo quality or ICSI outcome in women undergoing IVF/ICSI cycles. Accordingly, these biomarkers would be less reflective of the follicular environment and that the effect of maternal metabolic disorders on oocyte quality and fertility outcome is complex and of a multifactorial kind.
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).
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 : Although progress was made in an in vitro fertilization (IVF) tech niques, the majority of transferred embryos fail to implant. Morphology embryo scoring is the standard procedure for most of IVF centers for choosing the best embryo, but remains limited since even the embryos classified as (top quality) may not implant. Determining molecular mechanisms of human embryo implan tation is an extremely challenging task due to the limitation of materials and significant differences underlying this process among mammalian species. L-Se lectin and its ligand carbohydrate have been proposed as a system that mediates initial adhesion of human blastocysts to the uterine epithelia. Objectives: Quantitative evaluation of soluble L-Selectin molecule in an in-vitro culture media used for in vitro fertilization. Determine cut-off value of soluble L-Selectin quantity to be as qualitative predictor of successful implantation Patients and Methods: This prospective study was undertaken in the High In stitute of Infertility diagnosis and assisted Reproductive Technologies/ Al-Nah rain University/ Baghdad/ Iraq, during the period from September2017 to April 2018.A total of 74 infertile women were underwent controlled ovarian hyper stimulation for intracytoplasmic sperm injection cycle. Flexible antagonist pro tocol was used as ovulation induction protocol in all the cases .culture media L-Selectin level were measured on the day of embryo transfer by using Enzyme linked immune sorbent assay (ELISA) for all cases . Comparison in culture me dia L-Selectin was done to all cases. Results: There was no significant difference between pregnant and non-pregnant groups in age , BMI, and level of basal hormone FSH, LH , AMH and E2 in the day of hCG injection, and significant difference in the level of L-Selectin in pregnant compared to non-pregnant groups. Conclusion: Level of L-Selectin concentration in embryo culture media can be used as predictor of ICSI success
Background: The sperm plasma membrane activity and absence of antisperm antibodies in infertile patients ejaculate significantly associated with fertilization potential of human spermatozoa. The sperm viability detected by hypoosmotic swelling test (HOST) and autoimmune infertility detected by antisperm antibody assay presented diagnostic and prognostic tools for male infertility factors. Objective: This study was devised to compare the results of hypoosmotic swelling test and antisperm antibody assay for normozoospermic men and infertile patients affected by autoimmune infertility. Methods: Thirty semen samples were collected by masturbation and prepared by direct layering technique. Hypo-osmotic swelling test was performed by mixing 0.1 ml of semen with 0.9 ml of a 150 mOsm/L NaCl as a hypo-osmotic solution and direct immunobead assay were used to determine the presence of Antisperm Antibodies (ASA) bound on sperm surface. However, sperm concentration, sperm motility, and normal sperm morphology were evaluated according to World Health Organization (WHO) criteria and subjected to HOS test and ASA assay. The sperm prepared and incubated for 30 minute in 5% CO2 at 37°C after in vitro sperm preparation. Results: The results of the present study reported that the percentage of HOS test score in antisperm antibodies positive sperm samples (52.30 ±5.2) was significantly lower than that noticed in antisperm antibodies negative (70.30 ± 3.4) sperm samples. Conclusion: It was concluded that the use of HOS test as a simple and diagnostic and prognostic tools for identify male infertility. Further studies are suggested to assess the effect of ASA on sperm plasma functional integrity after ICSI and in vitro fertilization and embryo transfers (IVF-ET).
Background: Many studies has been found that the pentoxifylline (PX) and L-carnitine (LC) as motility stimulants showed a positive effect on the activation of sperm in vitro and improve the forward movement after cryopreservation. Objectives: The present study aim to investigate the possibility of using the motility stimulants(i.ePX,LC and mixture of PX with LC medium) for in vitro sperm activation after cryopreservation of epididymal sperms and their effects in vitro on fertilization rate (FR) and early embryonic development (ED) following in vitro fertilization (IVF) process as a model for obstructive azoospermia in men. Methods: Fifty mature male and Fifty mature female mice, 8-12 weeks old were used. Epididymal sperms were obtained from caudal region and in vitro direct activation technique was done after cryopreservation by using four media namely; PX alone, LC alone, a mixture of PX with LC and Ham's F-12 (as control media). The four media were used for IVF of mature ova and FR with ED were recorded. Results: In vitro activation of epididymal sperms with PX, LC and mixture of PX with LC has shown positive effect on sperm concentration, sperm motility, and grade activity of progressive forward movement after cryopreservation. There was a significant (P<0.05) increase of FR in treated media compared with the control medium after 24 hours of insemination. The study showed that the quality and quantity of embryos. Conclusion: It is concluded from the results of the present study that adding PX, LC and mixture of PX with LC medium to the culture medium of cryopreserved epididymal sperms and IVF leads to an improvement in certain sperm function parameters, supports the FR and early ED rate.
Placenta growth factor is, VEGF family member, a Pleiotropic Factor affects a variety of cell types and regulates various biological responses. PlGF is similar in structure and function to VEGF, and it amplifies VEGF's angiogenic actions. Ovulation stimulation during cycles of in vitro fertilization (IVF) for the management of infertility results in ovarian hyperstimulation syndrome (OHSS), an iatrogenic side effect. Considering potential roles of PlGF and its receptor (sflt-1) in angiogenesis, the association of these factors with OHSS among study women during controlled ovarian stimulation have evaluated in the present study. Comparative cross-sectional study including of 60 women who go through controlled ovarian stimulation. On oocyte retrieval day, their serum and follicular fluid were taken. The concentrations of PlGF and sFlt-1 were assessed using ELISA .Eighteen patients presented with ovarian hyperstimulation syndrome (OHSS) and 42 patients were no OHSS. There was significantly higher serum PIGF in women with OHSS patients as compared to women with no OHSS, 141.4 ± 11.8 versus 91.9 ± 5.4 respectively (p<0.001). Furthermore, Follicular fluid PlGF there was significantly higher in women with OHSS patients as compared to women with no OHSS, 163.4 ± 7.2 versus 91.1 ± 5.5 respectively (p<0.001). Additionally Follicular fluids PlGF/sFlt ratio significantly higher in women with OHSS patients as compared to women with no OHSS, 0.034 ± 0.01 versus 0.026 ± 0.01 respectively (p<0 . 001). On the contrary there was significantly lower serum and follicular fluid sFlt in ovarian hyper-stimulated patients These statistics show that serum and FF PlGF and PlGF to sFlt ratio higher in women with ovarian hyperstimulation syndrome (OHSS) patients as compared to women with no OHSS. So, the PlGF may be played an important a role in angiogenesis dysregulation.
The aim of any given ICSI cycle is to achieve normal response and avoid poor or hyper response. A part from ovarian hyper stimulation syndrome (OHSS), pregnancy rate in hyper responder females usually low which could be related to the adverse effect of hyper estrogenic state on oocytes, embryos quality and uterine endometrium. To evaluate the effect of hyper response on ICSI outcome represented by oocytes, embryos quality, fertilization and pregnancy rates. Study showed that total number of retrieved oocytes, mature, immature oocytes, total embryos, good quality embryos and blastocyst count were significantly higher in hyper followed by normal and poor responders respectively. Fertilization rate was significantly higher in both hyper and normal responders than poor ones, while pregnancy rate was significantly higher in hyper responders 47.6% followed by normal 40.7% and then poor ones 16.7%. Conclusions: Irrespective of abnormal hormonal milieu, excessive ovarian response and higher number of produced oocytes improve ICSI outcome in hyper responder females by increasing the chance of getting higher number of embryos, good quality embryos and even blastocyst numbers which in turn increase the chance of pregnancy.
Background: Since 1999 the World Health Organization (WHO) recommended the assessment of sperm morphology by Kruger strict criteria (KSC). The manual of WHO contains the cut off values of sperm morphology measured by KSC for normal subject and patients treated by in vitro fertilization (IVF) and intra- cytoplasmic sperm injection (ICSI) without any information regarding intrauterine insemination (IUI). Objectives: The objective of this study is to achieve two major goals that are:1- Assessment of sperm morphology by applying KSC, using two different biological stains, named commercially Testsimplets (TS)pre-stained slides and Sperm-O-Scan (SOS)stain 2- To have a cut off value for morphologically normal sperm that resulting into a successful IUI outcome. Materials and Methods: Fifty five unselected couples attending the Institute of Embryo Researches and Infertility Treatment were included in this prospective study from December 2007 till August 2008. The mean age of male partner was 31.7±1.87years with an average duration of 6.25±1.65years of infertility. The women were prepared for IUI. All the males were subjected to semen analysis with in vitro sperm activation using layering swim-up technique. Assessments of the morphologically normal sperm (MNS) of the samples were performed by using both, High power field and KSC by using TS and SOS stains before and after the activation. Correlation of the positive IUI results and MNS by KSC waw recorded denoting the cut off value for successful results. Results: The mean of morphologically normal sperm applying KSC with the use of SOS stain before in vitro sperm activation was 16.10% and after activation was 37.85% and by using TS stain before in vitro sperm activation was 15.14% and after activation was 37.09%. No significant (P>0.05) differences was detected between the results of MNS of the two stains. The result of IUI was 11 positive pregnancies out of the 55 cases. The mean of MNS using KSC for these positive cases was 11.58% and 12.84% for SOS and TS stains, respectively. Conclusions: The cut off value of MNS by applying KSC and using two different biological stains for a successful IUI program was 11.58% by SOS stain and 12.84% by TS pre-stained slides. Although the study found no statistical differences between the two stains, TS found to be easier, simpler and less time consuming technically than SOS stain.
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.
Endometriosis is a chronic inflammatory gynecological disease and a leading cause of female infertility, characterized by Th1/Th2 cytokine imbalance. Interleukin-13 (IL-13) and Interferon-gamma (IFN-gamma) are cytokines mediating anti-inflammatory and pro- inflammatory pathways, respectively, and were investigated in this study in infertile women with endometriosis undergoing Intracytoplasmic Sperm Injection (ICSI).To explore the potential impact and prognostic value of Interleukin-13 (IL-13) and Interferon- gamma (IFN-gamma) on ICSI outcome and to confirm their role in endometriosis-related infertility .A prospective case-control study was conducted involving 80 infertile women aged 18-40 years, including (n=40) women laparoscopically confirmed with endometriosis and (n=40) infertile women without endometriosis serving as controls. Both groups enrolled in ICSI cycles, and on the day of oocyte retrieval, serum and follicular fluid were collected; then concentrations of IFN-γ and IL-13 were quantified using enzyme-linked immunosorbent assay (ELISA). Ovarian stimulation characteristics, oocyte maturity, embryo quality, maturation rate, fertilization rate, and pregnancy rate were compared between groups using appropriate statistical analyses to investigate their association with endometriosis and their impact on ICSI outcomes. Elevated IFN-γ and IL-13 levels in both serum and follicular fluid are strongly associated with impaired ovarian response and poor ICSI outcomes in women with endometriosis. These findings suggest that inflammatory cytokine imbalance contributes to reduced oocyte developmental competence and embryo quality. Assessment of IFN-γ and IL-13 may provide clinically valuable biomarkers for predicting ART outcomes and could facilitate individualized therapeutic strategies targeting immune-mediated reproductive dysfunction.
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.
Infertility is the inability of a sexually active non-contraceptive couple to achieve pregnancy in one year. Sperm motility is one of the most important prerequisites to achieving fertilization and pregnancy . The objective of the current study was to improve certain sperm function parameters following in vitro activation of men suffering from asthenozoospermia by using a novel combination of motility stimulant substances, namely Maca, L-carnitine, and Pentoxifylline, to be used for assisted reproductive technologies programs in the future.Ninety asthenozoospermic patients were involved in the current study. Each sample was divided into two equal portions. One part was considered as a control by using Ham’s F12 medium, and the other part was considered as a treated group by adding medium content, a mixed combination of Maca powder extracts (Lepidium meyenii) (M)1mg/ml, Pentoxifylline (PTX) 10 mg /ml, and L-Carnitine (LC) 0.5mg /. Certain sperm function parameters were examined before and following in vitro preparation using the swim-up technique according to WHO guidelines.The results showed a significant (p<0.001) elevation in active sperm motility grade A, B, and progressive motility following in vitro preparation by Ham’s F12 media and mixed culture medium compared to before activation. The morphologically normal sperm percentage was significantly higher in mixed culture medium compared to control medium, and before activationWhile the results of DNA normality by using mixed culture and Ham’s F12 media were statistically shown a significant (p<0.001) reduction compared with before activation . It is concluded that the combination medium have a superior effects on certain sperm function parameters and DNA normality enhance its usage in sperm activation by swim up technique for ART programs.
Background: Gestational trophoblastic disease is a wide spectrum of abnormal pregnancy that includes: Complete molar pregnancy (CMP), partial molar pregnancies (PMP), invasive mole (IM), chorioncarcinoma (CHC), and placental site trophoblastic tumors (PSTT. Hydatidiform mole is an abnormal gestation characterized by the presence of hydropic changes affecting some or all of the placental villi. Hydatidiform mole arises as a result of the abnormal fertilization of an ovum. Tubal ectopic hydatidiform mole is a rare lesion and few cases have been reported.
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.
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.
Azoospermic patients have the option of undergoing in vitro fertilization procedures, specifically intracytoplasmic sperm injection (ICSI), for which sperm is retrieved from the patient's own testicles and then injected directly into the oocyte. The aim of the study: evaluate the role of repeated sperm extraction by enzymatic and mechanical mincing techniques in azoospemic patients. A total of 32 males with non-obstructive azoospermia participated in the research. Testicular sperm extraction (TESE) was performed under spinal or general anesthesia after a comprehensive history, physical examination, and investigations. After that, the biopsies underwent through a series of mincing processes, both mechanical and enzymatic. Patients gave information including age, body mass index and previous biopsy attempts. In both enzymatic and mechanical methods there was no significant higher sperm recovery in patients with history of previous testicular biopsies (66.7% vs. 38.5%; p=0.209) and (66.7% vs. 30.8%; p=0.102) respectively. Patients who had undergone prior testicular biopsies had marginally higher sperm recovery rates using both enzymatic and mechanical techniques
Background: Unlike sperm production in men which is continuous, women are born with all their eggs and they do not produce any more. Technique to bank eggs would allow girls and women to have the same reproductive options as men when faced with a serious disease and the long term complications of chemotherapies that threatens to destroy their eggs. Objective: The aim of this study is to improve the efficiency of existing ovarian cryopreservation, by following the most recent method named needle immersed vitrification (NIV) beside the slow freezing method. In this review we discussed also the historical description of the methods used, freezing and vitrification of mature and immature oocytes, this include comparison of principles, procedures and results as reported in the literature. Materials and Methods: A total of 285 fragments of human ovarian tissues were isolated and cryopreserved from 36 women aging 26 - 55 years old in the Reproductive Unit, Lubeck-Germany. Ovarian tissue strips of 1-3 mm thick of ovarian cortical biopsies obtained from patients. The collected human ovarian cortex fragments were assigned to two different cryopreservation procedures, slow-freezing and NIV. On the other hand, a total of 80 patients (100 cycles) with polycystic ovaries (PCO) were underwent in vitro maturation (IVM) treatment. The number of inspirated immature oocytes was 536. Vitrification of immature oocytes were carried out. Results: Successful freezing and storing were performed on 285 fragments of ovarian cortex tissue from 36 patients by two cryopreservation protocols as mentioned before but these tissues do not thaw and use yet. Concerning the IVM, the percentage of maturation, fertilization, survival, embryo transfer, and pregnancy rates were 55.2%, 44.5%, 96.2%, 88.1%, and 10.6%, respectively. Conclusion: Recent advances in freezing technology, modifications of conventional protocols used and continuing optimization of vitrification have efficiently improved the method. These results may lead to increase the possibility of preserving fertility by cryopreservation of ovarian tissue. Ovarian tissue banking can offer hope for cancer patients who want to safe guard their fertility against sterilizing chemotherapy and radiotherapy. Immature oocytes can be successfully isolated from the oophorectomy specimen regardless of the day of menstrual cycle, and undergo IVM and cryopreservation.
The effect of maternal body mass index (BMI) on fertility outcomes in women undergoing in vitro fertilization/intracytoplasmic sperm injection cycles has been extensively evaluated and the results of these studies have shown a lot of controversial issues. Folate is a naturally occurring type of vitamin B9 crucial for reproductive health. 65 infertile couples were subjected to intracytoplasmic sperm injection cycles. Both primary and secondary types of infertility were involved, with different causes. The mean plasma folate of all infertile women was 12.71±6.52, with pregnant 11.60±5.57 and non-pregnant 11.74±8.80; with no significant difference in mean plasma folate between them. Moreover, the means of follicular fluid folate of all infertile women, pregnant women, and non-pregnant women were 8.00±5.39, 7.84±4.68, and 8.39±6.19 respectively. There was no significant statistical difference in mean follicular fluid folate between pregnant and non-pregnant women (p=0.719). Also, both plasma folate and follicular fluid folate were not significantly correlated to oocyte and embryo characteristics. Although plasma folate was higher in obese than normal and overweight women, the difference did not reach statistical significance. It appears that the correlation among maternal BMI, folate level and fertility outcomes in women undergoing intracytoplasmic sperm injection cycles are still controversial and much research work is needed to figure out such complex interaction among these variables.
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.
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
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.
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 .
Background: Pretreatment of the semen samples of the asthenozoospermia patients with certain sperm stimulant agents prior to use for assisted reproductive techniques (ART) has been reported to have beneficial effects on fertilization rates. Objectives: Investigating the influence of addition of different concentrations of oxytocin (OT) to semen samples of asthenozoospermic patients for in vitro sperm activation as a new method for stimulation of sperm motility in these patients and exploring the concentration of OT that harvests amaximal numberofmotile sperms. Methods: Simple layer technique for in vitro sperm activation using culture medium enriched with OT was studied in asthenozoospermic patients (No. 90). Semen samples were examined before sperm activation, and then they were divided into three equal groups. In the first group, three concentrations of OT (2, 4, and 6 IU/ml) were applied. Based on the results of the mentioned work, OTat the concentration of 2 IU/mlwas applied with sperm preparation medium (SPM) and without in the second and third groups, respectively. Results: Results showed that there was a significantimprovementin the percentageof sperm motilityand spermgrade activity of the OT-treated samples atthe concentration of 2IU/mlas comparedto the control samples. Moreover, there was a tendency for a decrease in sperm grade activity with increase OT concentration. The outcome of sperm activation in presence of SPM revealed a significant increase in the sperm motility characteristics (the percentage of sperm motility and sperm grade activity) in the control and OT-treated samples and the best improvement in these sperm motility characteristics was observed in the OT-treated samples. When SPM was absent, a significant improvement in the sperm motility characteristics was observed only in the OTtreated samples. Conclusion: The results of this study indicate that OT at the concentration of 2 IU/ml has got a stimulatory effecton sperm motility characteristics; also, the use of SPM succeeds to supportthis effect.
Stem cell factor (SCF) is one of the first growth factors derived from granulosa cells in the ovarian follicle plays a critical role in hematopoiesis and the generation of melanocytes and germ cells. SCF also serves an important role in the development of the interstitial cells of Cajal in the intestine and the learning functions in the hippocampal region of the brain. This study aimed to investigate the levels of SCF in follicular fluid (FF) and if it can be used as a potential marker for predicting oocyte, embryo quality, and pregnancy rate. A total of 44 infertile couples involved in the study. All of them underwent controlled ovarian hyperstimulation for the ICSI cycle. Antagonist protocol was used as an ovulation induction protocol. Patients were divided into two groups according to ICSI outcomes as pregnant (23 women) and non-pregnant (21 women) groups. A comparison was done in both groups based on the oocyte, embryo quality, and other ICSI cycle characteristics, and follicular fluid stem cell factor level. SCF level was measured by using an enzyme-linked immunosorbent assay. Our study showed lack of association between patients' characteristics which included (age, BMI, infertility duration, type of infertility, and causes of infertility, basal hormonal, stimulation characteristics levels), FF SCF level and the oocyte characteristics comparison (total number retrieved oocyte, germinal vesicle oocyte (GV), MI oocyte and MII oocytes, and the number of injected oocytes, fertilization rate, and embryo characteristics) between pregnant and non-pregnant groups. The present study showed that Mean follicular fluid SCF was significantly lower in pregnant women in comparison with that of non-pregnant women, 135.92±38.96 pg/ml versus 185.15±63.58 pg/ml, respectively (P=0.003). Raised SCF level in FF harms ICSI outcome.
Mitochondria play a significant key in sperm metabolism as well as power generation. The mitochondria nucleolus is essential for several sperm activities. such as sperm capping, hyperactivation, and acrosome reactions, and oocyte fusion, mitochondrial membrane potential can be used to study sperm mitochondrial activity, the mitochondrial integrity of sperm is evaluated by examination of mitochondrial membrane potential MMP, which may be achieved by using fluorescent dyes like JC-1 probe is composed of 5,5,6,6-tetrachloro-1,1,3,3-tcarbocyanine iodide. To study the correlation between mitochondrial membrane potential Red MMP and sperm motility . This study included 42 male samples who were separated into two groups. (21 normozoospermic, 21 asthenozoospermia) through the period of attendance at the infertility clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, and Um Al-Banin Center for Management and in vitro fertilization the collected semen was obtained and seminal fluid analysis was assisted according to (WHO,2021). fluorescent staining of the sample prepared is done by using an MMP Assay Kit (with JC-1) was performed. A positive significant correlation between sperm motility percentage and red MMP intensity (p < 0.001). Mitochondrial activity plays an essential function in sperm motility and male fertility Male infertility, Sperm Motility, Mitochondrial Membrane Potential, MMP Assay Kit (with JC-1).
Background: Recently the reproductive hormones, Anti-Mullerian hormone (AMH), activin A, and follistatin (FS) have been found to play an important role in folliculogenesis, oocyte maturation and corpus luteum function by changing the pattern of granulosa cell expression which in turn affects the success of fertilization potential. Objectives: To examine the relation of in vivo concentrations of AMH, activin A, and FS hormones on the ovaries status in the phases of menstrual cycle and to elucidate the relationship of these hormones with pregnancy rate following intrauterine insemination(IUI). Materials and methods: Seventy seven infertile couples were recruited from infertility clinic population at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies through the period from September 2011 to May 2012. Those infertile couples were divided to 3 groups according to the infertility cause (male factors causes, anovulatory causes and unexplained causes of infertility). Measurements of AMH, activin A, and FS hormones levels were done at cycle day (CD25-). Then measurements of activin A, and FS hormones levels were done only at preovulatory cycle(CD1214-) when the size of the Graffian follicle range between 1824-mm and at least one follicle was present and before hCG injection and at day 28 of the cycle (CD28). IUI was performed 3640- hours after hCG injection then measurement of the three hormones was performed after successful IUI. The result of successful pregnancy was recorded and statistical relation with AMH, activin A and FS hormones was assessed. Results: There was a positive significant correlation between endometrial thickness (ET) and pregnancy rate and the best ET to predict pregnancy is more than 7mm. There was no significant relation (P>0.05) between AMH level and pregnancy rate. A significant correlation was found between follistatin levels in preovulatory (CD 1214-) with a cutoff value 0.20ng/ml to predict pregnancy. There was a significant relation between activin-A level and pregnancy rate with a predictive cut off value 397.5 ng/ml on (CD 28) only. Conclusions: Activin-A can be regarded as a biomarker candidate for diagnosing very early pregnancy at luteal phase following IUI. Follistatin preovulatory (average CD13) has the most predictive value for pregnancy following IUI.
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.
Background: Glycyrrhiza glabra(Gg) is a herbal medicine used in the treatment of different syndromes. Recently, the effect of this plant on the reproductive system has been studied. However, its role in in vitro still not clear. Objectives: The present study aims to investigate the possibility of using Glycyrrhiza glabra extract for in vitro sperm direct activation, and its effect on in vivo fertilization rate (FR) and early embryonic development (ED) following intra-peritoneal insemination (IPI). The mice were used as an experimental model for mammals. Materials and Methods: Glycyrrhiza glabra extract (2 mg/ml culture medium) was used for in vitro direct sperm activation technique. The female mice were divided into two groups: spontaneously ovulated (SOM) and superovulated groups (SuOM). Each group was divided into two subgroups, the first: the IPI was accomplished by epididymal sperm activated in vitro by adding 20%Gg to the culture medium. The second subgroup: the IPI is accomplished without adding Gg to the culture medium. Results: In vitro activation of epididymal sperms with 20% Gg has shown positive effects on sperm concentration, sperm motility, and grade activity of progressive forward movement. There was a significant (P<0.05) increase in FR with adding 20%Gg compared with that free of Gg of spontaneously ovulated groups after 24 hours of insemination. The FR in the right oviduct showed highly significant (P<0.001) increment compared to the left. Embryonic development rate significantly ( (P<0.05) increased after 24 and 48 hours of insemination in grouping with adding Gg. The study showed that the quantity and quality of embryos generated from IPI with adding Gg were higher than that of groups inseminated without adding Gg. Conclusion: It is concluded from the results of the present study that adding the 20% Gg to the culture medium of the epididymal sperm and IPI leads to an improvement in certain sperm function parameters and supports the FR in SOM and SuOM with an increase in the embryonic development rate. These data can be utilized for artificial insemination programs.
During the process of embryo development, endometrial receptivity is very important for the successful implantation of blastocysts. This research investigated the influence of endometrial thickness on the day of oocyte retrieval and fresh embryo transfer on pregnancy outcomes in Iraqi women who are undergoing intracytoplasmic sperm injection (ICSI). Women from Iraq are taking part in this investigation. Participating in in vitro fertilization (IVF) cycles with the use of ICSI was a group of 44 women who were diagnosed with polycystic ovarian syndrome (PCOS) and had unexplained infertility. During the phases of oocyte retrieval and embryo transfer, there were significant differences seen between the two groups (p-values of 0.003 and 0.049), respectively. There was a total of 44 individuals, and 31 of them had negative pregnancy test results. The remaining 13 persons had positive findings. These results illustrate the importance of endometrial thickness in influencing the effectiveness of assisted reproductive technologies in achieving pregnancies.
Women who have transvaginal oocyte retrieval feel nervous and have mild to moderate pain because a needle is used to poke holes in the vaginal skin and ovarian capsule to get the oocytes. The aim of this study is compare between general and spinal anesthesia effect on ICSI outcome . A cross-sectional study was conducted in the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies/ Al-Nahrain University, from first April 2021 to the first of February 2022. Patients were randomized blindly at time of oocyte retrieval for types of anesthesia given: (50 women subjected to spinal anesthesia and 50 women subjected to general anesthesia). All females were subjected to flexible antagonist protocol. Also, all of them had the same surgical procedure by the same physicians. ICSI outcome was compared between the groups. The study showed there was non-significant difference (p>0.05) between the two groups regarding pregnancy rate with higher oocyte number in women subjected to spinal anesthesia than those who were under the general anesthesia although the difference was not statistically significant. The fertilized oocyte number was non significantly higher in spinal anesthesia group than the general anesthesia. So; spinal anesthesia and general anesthesia have comparable ICSI outcome.
Background: Semen quality is a measure of the ability of semen to accomplish fertilization. Semen quality involves both sperm quantity and quality. The general population is exposed to metals at low concentrations either voluntarily through supplementation or involuntarily through intake of contaminated food and water or contact with contaminated soil, dust, or air. Some metals, such as lead (Pb), are nonessential xenobiotics that can be measured in most of the general population. Objectives: Our goal in this study is to assess relationships between exposure to lead at environmental levels and human semen-quality parameters. Materials and Methods: The study populations consist of 81 infertile male of general population who are occupationally unexposed to lead and another 26 healthy fertile male as control group. Blood lead and seminal plasma lead measurements performed by the electrothermal-atomic absorption spectrometry (AAS) method. Macroscopic and microscopic examination of semen performed according to WHO recommendations. Seminal plasma separated from the spermatozoa by centrifugation and stored at -20°C until required for analyses. Results: This study showed significant increase in seminal plasma lead mean value in moderate and severe oligozoospermic infertile males than in fertile male controls( p<0.05). The result also showed no statistically significant difference in blood lead concentration among the infertile group compared to fertile controls. Conclusions: The results of this study indicate that even moderate exposure to Pb at environmental level (blood Pb < 400 µg/L) can significantly reduce reproductive capacity in men.
The vaginal microbial makeup has a substantial impact on women's health. The typical vaginal microbial profile in healthy women of reproductive age comprises aerobic, facultatively anaerobic, as well as obligately anaerobic organisms. However, the optimal vaginal microbiota is dominated by lactobacilli and is thought to defend against pathogenic microbial colonization and infection by producing lactic acid, antimicrobial metabolites, and lowering immune system activation. Recent research has come up with the term "molecular bacterial vaginosis (BV)" to describe non-optimal vaginal microbiota that is low in Lactobacillus species (spp.) and has a lot of facultative and absolute anaerobic bacteria specifically Gardnerella vaginalis. In the context of IVF, BV has been linked to infertility. The study comprised forty-six women to explore the vaginal and follicular microbiota of asymptomatic infertile Iraqi women starting an ICSI trial. At the time of ova pick-up, a high vaginal swab and follicular fluid were acquired from each instance for DNA extraction to test for the presence of microorganisms by the use of a real-time PCR special commercial kit. In conclusion, molecular technologies have offered a more thorough view into the vaginal microbiota, which may explain why vaginal dysbiosis shifts during the ICSI-ET treatment. Even though the majority of women have no clinically visible infection, the fact is that they have an aberrant vaginal micro-ecology profile. Nevertheless, isolating microorganisms from follicular fluid did not reduce the likelihood of fertilization or the pregnancy rate during ICSI cycles.
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.