This study was designed to assess the effect of different concentrations of L-carnitine and Co enzyme Q10 (COQ10) supplied to SMART Pro-medium on human sperm motility and sperm grade activity ( progressive, non-progressive and immotile) during in vitro sperm activation. Eighty seven samples of semen of infertile and fertile men were randomly collected from the subjects at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies / Al-Nahrain University. The mean age of the men was (32.034 ± 0.57) years with mean duration of infertility (4.644 ± 0.25) years. Seminal fluid analyses were done involving macroscopic and microscopic examinations according to WHO criteria. Swim-up technique was dependent for in vitro sperm activation , the washed samples were divided before using centrifugation swim-up technique into 3 groups: control group (G1) without L-carnitine and CoQ10. While, in G2, G3 two concentrations of L-carnitine and CoQ10 (20 µg , and 40 µg ) were used. Moreover, all groups within post-activation enriched with SMART-Pro media. The sperm motility and sperm grade activity were evaluated after the addition of L-carnitine and COQ10. The results showed a significant (P<0.05) differences were showed in sperm motility and sperm grade activity of post-activation when compared with the pre-activation. In G2 and G3, with dose of ( 20 µg and 40 µg) of L-carnitine and CoQ10, the results showed a significant (P<0.05) increase in the percentages of sperm motility, progressive sperm motility in spite of the increase in the percentages of sperm motility, progressive sperm motility in G3 compared to G2, there was non significant (P>0.05) differences between G2 and G3. The results for non-progressive sperm motility% and total progressive sperm motility millions/ mL. for 40 µg ( G3) groups were significantly (P< 0.05) decreased when compared to control group (G1) with non-significant (P>0.05) differences between 20µg (G2) group, with significant (P< 0.05) decreased in immotile sperm percentage in post-activation when compared to the pre-activation. From the results of the study, it can be concluded that the addition of 40 µg of L-carnitine and CoQ10 to washed sperms can improve sperm motility and sperm grade activity in vitro. Data were analyzed statistically using complete randomized design (CRD) (one way ANOVA).
Background: Semen preparation techniques were developed to separate motile sperm that are morphologically normal from seminal plasma to optimize successful assisted reproductive technology cycles which seem to be the most effective options in cases of a male factor infertility. Objective: The objective of the present study was to compare between outcomes of two sperm preparation techniques for asthenozoospermic patients includes:1-Centrifugation swim up technique.2-Glass wool filtration technique. Subjects, Materials and Methods: Fifty three infertile males were participated in this study during their attendance to the Infertility Clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies; Al-Nahrain University. Semen samples were collected and SFA was done according to WHO (2010 and 1999). Each semen sample was divided into two aliquots. The first one prepared using centrifugation swim up technique, while the other one prepared using glass wool filtration technique then sperm parameters were assessed for both techniques and the results were statistically analyzed. Results: After in vitro sperm activation using both techniques, there was significant reduction (P<0.05) in the sperm concentration, significant improvement (P<0.05) in the percentages of sperm motility and morphologically normal sperm when compared to pre-activation. Present study appeared that the glass wool filtration technique resulted in significantly (P<0.05) better results for sperm concentration and total number of progressive motile sperm than the swim-up technique. The present study proved that there was significant (P<0.05) improvement in sperm parameters (increment for sperm motility (%), progressive sperm motility (%), total number of progressive motile sperm and normal sperm morphology (%), while reduction for round cell count and sperm agglutination percent) for all cases using glass wool filtration techniques. In contrast, there was failure of sperm activation for 10 cases using centrifugation swim up technique. Conclusions: From results of the present study, the sperm parameters outcomes using glass wool filtration technique was superior to the outcomes of centrifugation swim up technique when prepare semen of asthenozoospermic patients.
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.
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
Background: There are many factors and media affecting sperm characters which are defined by classical parameters namely, concentration, motility and morphology, according to standard World Health Organization (WHO). Objectives: To study the effect of incubation time on certain sperm function parameters following in vitro centrifugation technique by sperm wash, FertiCult medium for the purpose of assisted reproductive techniques (ART) in couples presenting sub-fertility. Methods: Two groups of 40 semen samples (group 1: fertile men with normal semen parameters and group 2: abnormal semen parameters) underwent sperm activation test (SAT) in the ART laboratory, High Institute of Infertility Diagnosis & ART/Al-Nahrain University. Semen analysis was assessed in standard WHO (1999). Wash and spin activation technique by FertiCultTM medium was done for all samples. The tubes which was sealed, inclined vertically and kept at 37°C for 30 minute. The supernatant contained actively motile sperms was transferred to another test tube to separate that active sperm from pellet and record sperm parameters after. Results: There is a significant (p<0.05) improvement in certain sperm parameters after 30min. of incubation following SAT in both groups compared to before activation and following 30min. incubation in SAT. Certain sperm characters are significantly decline in group 2 after 1 hr. incubation compared to before activation. Conclusions: Sperm activation test is very important method to perform before undergone artificial insemination (AIH). Incubation of prepared sperm in vitro by sperm wash, FertiCult medium is benefit for the first half hour to all semen samples to have a success in artificial insemination.
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.
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.
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.
Background: Sperm cells are well equipped with a powerful defense system of antioxidants, but an imbalance between the production of reactive oxygen species (ROS) and the available antioxidant-defenses result in oxidative stress. Therefore, antioxidants are supplemented extracellularly under in vitro conditions. Antioxidants are the main defense factors against oxidative stress induced by free radicals. Vitamin E is believed to be the primary component of the antioxidant system of the spermatozoa and is one of the major membrane protectants against ROS and lipid peroxidation (LPO) attack. It appears to be the first line of defense against the peroxidation of polyunsaturated fatty acids (PUFAs) contained in the cellular and sub-cellular membrane phospholipids because of its lipid solubility. Objective: The aim of the current study to evaluate effects of supplementing semen extender with vitamin E, at various concentrations (2, 6 and 12 IU/ml) on sperm motility, survival and morphology on the releases of free radicals and antioxidant enzymes within semen. Patients ,Materials and Methods: Twenty two patients were involved in the present study semen samples were obtained, and sperm count was assessed. The antioxidant (vitamin E) was formulated to be tested at three different levels as follow; 2, 6 and 12 IU/ml; Therefore, control and 3 antioxidant-containing extenders were prepared for semen dilution. The sample was divided into 4 aliquots. One volume of semen was added to 5 equal volumes of the designed extender. Extended semen samples (37˚C) were gradually cooled to 4˚C in the refrigerator and stored for 48 hours. A semen sample was taken out after 48h of storage, then warmed to 37˚C and checked for progressive motility, viability and abnormality. Sperm viability was assessed by Eosin Y. (0.5%)—Nigrosin (0.1%) staining mixture. A total of 200 sperm were assessed under oil immersion with a high-resolution (X100) objective. Sperm morphology was assessed by Hematoxylin-Eosin satin, at least 200 sperm were scored on randomly chosen field, under oil immersion with a high-resolution (X100) objective. Results: The highest post-thaw motility and sperm survival (44% and 51.7%) 48 h after storage at 4˚C (Table 1) were attained (P < 0.01) in the basal diluents containing 6 IU vitamin E. Likewise, the correspondent percentages of live spermatozoa were 59%. Contrariwise, percentage of sperm abnormalities for the previous treatment was 25% significantly (P < 0.05) decrease compares with basal diluents containing 2, 12 IU vitamin E. The lowest (P ≥ 0.05) survival (23.5%, Table 1) was found in the basal diluents containing 12 IU vitamin. Beyond 6 IU vitamin E, there appears to exert adverse effects on sperm viability and survival. Conclusions: For the best protection against the increased free radicals production during chilling-preservation of human semen, extenders must contain 6 IU vitamin E per ml. The intensive production of free radicals in seminal plasma was counteracted by the inclusion of vitamin E at the above mentioned concentrations.
Background: All men have the natural desire to have their own biological children. Cryopreservation of male gametes is an important aspect of human fertility preservation. With the advancement in assisted reproductive technology, indication for sperm cryopreservation is expanding. Objectives: To evaluate the effect of cryopreservation by using Sperm Freeze TM kit on some sperm parameters for infertile patients. Materials and methods: Ninety five semen samples were taken from male patients. The seminal fluid analysis was performed according to the standard criteria of WHO (1999). The patients were divided into four groups depending on their sperm concentration and motility: Normozoospermic, oligozoospermic, asthenozoospermic and oligoasthenozoospermic patients. Cryopreservation in liquid nitrogen was done by using Sperm Freeze TM kit for the samples and thawing was performed after three months. Results: In general, the post-thawing study revealed a considerable highly significant (P <0.001) decrease in the sperm parameters which include the means of sperm concentration, sperm motility, progressive motility and normal sperm morphology as compared to that of pre-cryopreservation. Whereas the percentage of immotile sperm was highly significant (p<0.05) increase as compared to pre-cryopreservation in all groups except for oligozoospermic patients in which there was a significant (p<0.05) increase. Conclusions: After semen cryopreservation, all parameters were significantly decreased.
Human sperm chromatin and integrity of DNA have multiple endogenous factors, such as sperm compression and nucleoproteins, with various mechanisms all through their own sperm production and transport. The integrity of the paternal genetic material in both regular and assisted pregnancy is essential for initiating and maintaining a viable pregnancy. The study included semen samples from 100 men taken from Al Nahrain University Medical Clinic and Babylon Teaching Hospital for Women and Children. The first sperm test was based on the World Health Organization (1999), followed by a DNA integrity test that uses a DNA fragmentation test, and the third intensification test Chromatin by aniline blue dye. It was correlated negatively with the sperm count in combination with DFI and chromatin sperm with semen characteristics (r = - 0.223, p = 0.026). As well as DFI, sperm chromatin was being clinically significant associated with the rate of round cells (r = 0.39, < 0.001). DFI was positive and significantly associated with sperm chromatin in all infertile men (r = 0.958; p<0.001). DNA isolation in all enrolled men has been adversely linked with sperm/ejaculation, has been associated with the round cell count positive way and is not understanding and resolving with other parameters. The width of abstinence and beneficial cell count in all register men were strongly linked with Sperm chromatin. There was a direct relationship between the fragmentation of DNA and chromatin. This could imply that one exam can be used instead of the other.
Background: The effect of L-carnitine (LC) on the male infertility have been previously reviewed. However there are few studies about the effect of LC on sperm function parameters in vitro. Objectives: The objective of this study was to improve certain sperm function parameters in vitro of asthenozoospermic infertile men using LC as a motility stimulant substance which may enhance the medium used for assisted reproductive technologies (such as artificial insemination) in future. Methods: Semen was collected from 50 infertile men involved in the current study. Each semen sample was divided into two portions. One part was considered as a control and in vitro activated by using culture medium only. The other portion was considered as treated group and in vitro activated by adding LC (0.5mg) to the culture media. Certain sperm function parameters were examined before and following in vitro activation using simple layer technique. Results: The results revealed highly significant increment (P<0.01) in the percentage of total sperm motility grades (A+B+C) and progressive sperm motility grade (A) with a significant improvement (P<0.05) in the percentage of progressive sperm motility grade (B) and morphologically normal sperm when using LC medium in comparison with control medium after 30 minutes incubation. Conclusion: It is concluded from the results of the present study that adding 20% LC to the culture medium ofthe in vitro sperm activation leads to an improvement in certain spermfunction parameters.
Background: White blood cell are present in most human ejaculates, but leukocytospermia may reflect an underlying pathological condition. This study evaluated the relationship between leukocytospermia and standard semen parameters. Objectives: To investigate the presence of correlation between the leukocytospermia, sperm motility and sperm concentration in infertile Iraqi men. Methods: A total of 154 semen analyses of infertile men were included in this study. All semen analysis were performed within 1 hour, Data were collected for sperm motility type A and B, concentration of both leukocyte and spermatozoa. The mean numbers of motile sperms and the leukocytes were obtained for each specimen. 10 micro liter of seminal fluid were transferred on a slid and covered by a cover slip, then the mean number of motile sperm, leukocyte and concentration were obtained by reading 25 Microscopic fields. Results: The results indicated a significant relationship between the reduction in the number of leukocytes and the increase in the number of motile spermatozoa and sperm concentration r = -0.731 and r = - 0.77 respectively. But the mean number of motile sperms was weakly correlated with the sperm concentration r = 0.58. Conclusion: Both the mean number of motile sperms and sperm concentration were significantly correlated with the increasing number of leukocytes. That refers to improvement in motile sperm and sperm concentration with the reduction of leukocytes.
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.
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).
The objective of the research was to investigate the potential correlation between serum anti-Müllerian hormone (AMH) levels and sperm parameters such as sperm concentration, sperm count, sperm motility, and sperm morphology. A hundred male patients voluntarily participated in the study at a private andrology clinic in Al Najaf Al Ashraf City, Iraq, between October 2023 and November 2023. The patients were divided into four groups: 26 patients with oligozoospermia, 21 patients with asthenozoospermia, 28 patients with oligoasthenoteratozoospermia (OAT), and those with normozoospermia. Semen quality was evaluated by conducting a semen analysis according to the 2021 World Health Organization guidelines. The ELISA method was employed to measure the levels of AMH in serum samples. Biotechnology Instrument, USA, manufactured the ELIZA components that were used. The average age of the participants was 30.85 ± 0.70 years, BMI was 27.60 ± 0.58 kg/m2, and serum AMH was 2.08 ± 0.11 ng/ml. We did not find any negative or positive correlation between AMH levels in the blood with sperm concentration, sperm count, sperm motility, and sperm morphology in all study groups. The results of this study showed that serum AMH is an unreliable marker for predicting spermatogenic abnormalities.
ackground: Pretreatment of semen samples of the infertile patients and in vitro preparation techniques with the certain sperm stimulant agents prior to use for assisted reproductive techniques (ART) has been reported to have beneficial effects on pregnancy rates. Objectives: The objective of this study is to improve certain sperm function parameters in vitro of infertile patients semen by using novel combination motility stimulants substances that improved medium to be used for ART in future. Patients and Methods: One hundred infertile men involved in the current study. They were divided into four groups. Each semen sample was divided into four portions. One part was considered as a control and in vitro activated by using culture medium only. The other three portions were considered as treated groups and activated in vitro by adding pentoxifylline(PX,1mg/ml) and/or L-carnitine (LC,0.5mg/ml) to the culture media. Certain sperm function parameters were examined before and following in vitro activation using layering and sedimentation techniques. Results: The results showed a highly significant (P<0.001) improvement in the sperm concentration and percentage of progressive sperm motility grade(A), grade (B) and grade (A+B) with a highly significant (P<0.001) increase in the percentage of morphologically normal sperm (MNS) after addition of PX and/or LC to the culture medium in comparison with control medium using layering and sedimentation(layering under paraffin) techniques in the infertile patients. Conclusion: The results of this study indicate that adding 20%PXand/ or LC to the culture medium for in vitro sperm activation using layering and sedimentation techniques leads to an improvement in certain sperm function parameters.
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).
Background: Seminal fructose is derived from the seminal vesicle and is therefore a suitable marker for its function. Fructose is the energy source for sperm and may have an importance in pathological conditions of semen, e.g. in asthenozoospermia. Objective: To find out if there is a correlation between seminal plasma fructose level and sperm concentration and motility in infertile Iraqi men. In addition, the study assessed the reliability of using seminal fructose concentration as indicator of seminal vesicles function. Patients and Methods: Forty six infertile men, free from infectious disease or endocrine disorders, were selected for the study. Control group, selected from couples with proven male fertility, were 10 fertile men. Semen samples, collected from theses fertile and infertile men, were analyzed for general seminal fluid examination and fructose concentration determination. Results: Seminal fructose has been noticed to have significant negative correlations with sperm count, with percentage of motile sperm and with absolute number of motile sperms. These negative correlations may be due to increased rate of fructolysis with higher sperm count and motility or due to an endocrine cause related to relatively high FSH in infertile men. To use seminal plasma fructose as a marker for seminal vesicles function, a correction for the opposing factors that affect fructose level such as sperm count and motility is carried out. Corrected seminal fructose is positively correlated with the total number of motile sperm. Furthermore, a positive correlation between true corrected fructose and the percentage as well as the total number of motile sperm has been found. Conclusion: Seminal fructose level has an inverse correlation with sperm count and percentage and number of motile sperm in the study group. Seminal fructose is a good marker of seminal vesicle function after using the correction formulae.
Background: Many plants have medical importance due to the presence an active compounds in plants extraction acting as a drugs for treatment of several diseases in human and animals. Euphorbia helioscopia extract was used to treat diabetes, removal the warts of the skin, stimulate the nervous system and also used against constipation. Objective: Study the effect of terpinoid extract of E.helioscopia on rat epididymal sperm activity and fertility efficacy of male rats. Methods: This study was conducted at Biol. Dept., College of Science/ University of Babylon and Medical College, Mahidol University- Thailand. The study was designed to purify the active compounds of the Terpinoid of Euphorbia helioscopia extract by Nuclear Magnetic Resonance (NMR), and studying their effects on sperm motility and grade activity of the epididymal sperms of the rats. Six main active compounds were isolated from the terpinoid extract of E. helioscopia due to the differences in color, weight and nature by using NMR. The main terpinoid compounds were named A,B,C,D,E and F. Two concentration (0.5 mg/ml and 1 mg/ml) of each active compound of E. helioscopia extract were added to sperm suspension in vitro and incubated for 30 minutes, then sperm motility and grade activity were assessed and the results were compared to those values before adding the active compounds (control). Results: The results showed that 0.5 mg/ml of C and D active terpinoid compounds caused significant (p<0.05) decrease in sperm motility compared to before adding the extract, while the same concentration of B,C,D,E and F active terpinoids compound caused a significant (p<0.05) decrease in grade activity. The addition of 1 mg/ml of terpinoid active compounds (C,D and F) caused a significant (p<0.05) decrease in sperm motility, while the grade activity significantly (p<0.05) decreased by adding B,C,D,E and F active terpinoid compounds compared to control. The sub - compounds (C1,C2,D1,D2,D3 and D4) had a powerful decreasing effect on the sperm motility and grade activity compared to control and other active compounds. Conclusion: It was concluded that terpinoid compounds of E. helioscopia extract have adverse effects on rat epididymal sperm activity.
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: Semen cryopreservation is a useful tool for preserving fertility for men involved in ART program who have been diagnosed with cancer and will undergo chemotherapy, radiotherapy or testicular surgery. Objective: The objective of this study to evaluate the effect of honey bee supplemented to cryoprotectant medium on post-thaw sperm DNA integrity ,morphology and round cells. Subjects ,Materials and Methods: Thirty semen samples were collected from 30 infertile patients. After assessment of semen analysis, semen samples were divided into 3 aliquots (0.7ml for each sample) and mixed with 1 ml of cryopreservation solution (G1,control) alone, enriched with 5% honey bee(G2) with 10% honey bee (G3) for cryopreservation. Current study used the acridine orange test to investigate the effect of honey bee in two concentration (5%, 10%) on the integrity of sperm chromatin structure pre-and post cryopreservation. Cryopreservation was done at -196 Co in liquid nitrogen and thawing was performed after six months. Direct swim up technique was used for in vitro sperm preparation post-thawing, sperm parameters were assessed and data were statistically analyzed pre-and post-thawing. Results: The results show there was a significant(P<0.05) increase in the percentage of morphologically normal sperm for all groups post-thawing, particularly for G3 group that was significantly (P<0.05) increased as compared to G1 and G2 groups, In contrast, non-significant differences (P>0.05) were observed between G1 and G2 groups. Sperm DNA fragmentation percentage (%) was significantly(P<0.05) increased in G1 post thawing without any additives as compared to pre-cryopreservation , where as in G2 and G3 sperm DNA fragmentation were increased but not significantly as post-cryopreservation groups. Also the results show that there were no significant differences between G2 and G3 whereas G3 gives better result as compared to G1 with significant differences. Round cells counts for all groups of post cryopreservation were significantly(P>0.05) decreased as compared to pre-cryopreservation. Non significant (P>0.05) differences in the round cells counts among all the groups of post -thawing in spite of G3 group has the lowest mean. Conclusions: From these results we concluded that supplementation of 10% of honey bee to the freezing cryoprotectant medium gives best protection to the morphology and DNA integrity of human sperm.
Background: 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).
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: 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: Obesity has been defined as abnormal or excessive fat accumulation that may impair health while the Body mass index (BMI) is a simple index of the weight-to-height ratio that is commonly used in classifying overweight and obesity in adult populations. Many semen parameters alteration can be ascribed to obesity like decreased sperm concentration, abnormal morphology, compromised chromatin integrity and abnormal motility. Many evidences showed an adverse effect of excess body fat on spermatogenesis. Methods: A total sample size composed of (70)infertile men were grouped into three groups according to published BMI ranges as follows: normal weight, overweight and obese.A single semen sample was collected for each infertile men.Heightin (m) and weight in (kg) were recorded on day of semen collection.BMI was calculated as BMI= Kg\m2. Sperm parameters evaluated according to World Health Organization guidelines.Quantitation of the IL-6 in seminal plasma were accomplished using commercially available ELISA kit. Results: In the current study, a positive correlation was found between body mass index and IL-6, on the contrary, a significant negative correlation was found among IL-6 level and progressive sperm motility and sperm concentration as well. Conclusion: There is an increase of IL-6 from the adipose tissue with increasing BMI. The IL-6 cause reduction of the sperm motility by increasing the level of ROS. The obesity reduces the spermatogenesis by androgenic deficiency results in low sperm concentration and motilitywhich it may be the main reason of their infertility.
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: 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.
When evaluating male reproductive potential, it is crucial to evaluate sperm structural abnormalities and sperm DNA damage. Apoptosis and mitochondrial membrane potential disruption are linked to sperm DNA fragmentation (SDF), which is inversely correlated with motility and normal morphology. To enable liquefaction, the semen sample is taken in a sterile collection container and incubated at 37°C for 30 minutes The samples were then split into three groups, P, G, and N, based on the morphological %. To measure chromatin condensation, aniline blue staining (AB), an acidic dye with a strong affinity for non-lysine-rich histones, was employed. The DNA maturity of the spermatozoa was assessed using AB. Sperm concentration, total sperm count, total motility, and total progressively motile sperm all had means that were above the WHO 2010 standard range. The DNA fragmentation index was significantly higher in patients with pattern P than in patients with patterns G and N, and it differed significantly (P=0.003) between the semen morphological patterns. In conclusion, as DNA fragmentation is an independent test this study showed that a low percentage of normal sperm morphology might indicate DNA integrity status.
The integrity of sperm deoxyribonucleic acid (DNA) and chromatin is very important for genetic material transmission into offspring. Therefore, the aim of the present study was to investigate the correlation between sperm deoxyribonucleic acid integrity and sperm parameters. The current study was conducted on 96 semen samples. This study was done at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, for the period between November 2020 and May 2021. The samples were collected, and seminal fluid analysis was performed according to World Health Organization guidelines. Sperm Chromatin Dispersion test was performed. The results showed that the deoxyribonucleic acid fragmentation of sperms in infertile men in this study was higher than reported in fertile men in previous studies, indicating deoxyribonucleic acid fragmentation (DNA) is actively contributing at least partially to male infertility. In terms of enrolled men, sperm deoxyribonucleic acid fragmentation (SDF) showed a significant positive correlation with round cell count.
For conception and the development of healthy embryos, sperm DNA integrity is crucial. According to a growing body of studies, there is a strong correlation between sperm DNA damage and male infertility. Among the new medicines being developed in the medical field, the application of Platelet Rich Plasma (PRP) in human reproduction has yet to be examined. A total of 100 semen samples were used in the current experimental investigation. From November 2020 to June 2021, the research was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Masturbation was used to get an ejaculated semen sample. After semen analysis, the samples were separated into two equal parts, one without autologous PRP and the other with 2% autologous PRP, with the DNA fragmentation assessed using the Sperm Chromatin Dispersion Test. There was highly significant reduction in DNA fragmentation index (p < 0.001). The mean sperm DNA integrity was reduced after adding PRP (33.85±16.73 vs 38.55±16.64), Mean (± SE). PRP has been shown to improve human sperm DNA integrity.
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.
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.
Background: Metformin is anti-hyperglycemic effect due to decrease of hepatic glucose production. Another effect, leading decrease insulin resistance, regarded a safe drug, treatment of choice for overweight. Objective: To investigate the effects of 3 concentration of MF administered orally to male mice on sperm parameters as model for human being. Materials and Methods: In current study, 80 male mice were taken with age ranging between 1.5-2 month and weights ranging between 25-30 grams males classified into four groups of a control group (G1) and treated group (G2, G3 and G4) doses of MF were employed, 0.2, 0.4, 0.6 mg/mice daily to groups, given orally for 6 weeks and then the animals were scarified. Results Results have revealed a significant difference (P < 0.05) in sperm concentration for 3 groups compared to G1 group. Progressive motility (%) appeared with significant difference (P< 0.05) for G3 group as compared to other treated and G1 group. Furthermore, the result illustrated a significant decreased (P< 0.05) in the abnormal sperm morphology for G3 group compared with treated and control. However, sperm agglutination (%) confirmed that the G3 group exhibited highly statistical significant decrease (p≤0.001) compared to G1, G2 and G4. Conclusions From results of the present study, it was concluded that the administration of MF doses reduce sperm concentration .while, 0.4mg dose improve certain sperm parameters.
Background: The increasing prevalence of infertility cases is becoming a major public health problem in developing countries due to changes in diet and lifestyle. Objectives: Black Carrot Extract BCE (Daucuscarota) plays an important role as fertility agent due to its antioxidant and anti-proliferative properties. Therefore, the aim of the present study was to identify the effects of different concentrations of BCE on the outcome of in vitro human sperm activation of infertile patients. Methods: Thirty two semen samples were prepared which served as a control, using direct swim-up technique. Modified Earl's medium (MEM) was used only in group 2 (G2) and two concentrations of BCE (10µg/1ml and 20µg/1ml) were added to culture media MEM of groups 3 (G3) and 4 (G4) for in vitro sperm activation (ISA). Results: A highly significant (P<0.001) correlation was noticed between G4 with G2 and G3 in progressive motility when activation by adding 20µg/1ml of BCE to culture media as compared to G3 (which adding 10µg/1ml of BCE to culture media) and G2 (activation with culture media only). Moreover, swim-up activation technique with culture media and by both adding (10µg/1ml and 20µg/1ml of BCE respectively) to culture media enhancement in the (sperm motility, progressive sperm activity and sperm morphology). Conclusion: The high concentration of BCE is effective in the enhancement of progressive sperm activity during in vitro human sperm activation.
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.
Intrauterine insemination (IUI), it is a type of artificial insemination, used to treat infertility by which the Sperm that have been prepared and placed directly into the woman’s uterus around the time of ovulation. The main purpose of this procedure is to enable the sperm to swim into the fallopian tubes and fertilize a ready egg in order to lead to a natural pregnancy. IUI can be coordinated with the normal cycle with fertility or medications depending on the reasons for infertility The IUI outcome still low about 20-25% so adding number of biochemical factors to the culture medium which are essential to regulate the fertility potential of spermatozoa, for example, platelet-activating factor (PAF) which is a unique and novel signaling phospholipid that has multiple biologic properties have been tried. To determine the effect of adding the PAF into the culture medium to prepare and activate the semen samples of non-male factor infertility (normal semen analyses). This study was carried out in the Infertility Unit, AL-Hussein Teaching Hospital, Al-Nassirya City, through the period from 30th of October 2020 till 17th of April 2021. The study included thirty couples who are complaining from infertility (they failed to conceive after one year at least of regular unprotected intercourse. The frequency distribution of enrolled infertile men with normozoospermia Baseline characteristics of seminal fluid analysis of males with normozoospermia those with PAF are Semen volume ml, pH, Sperm concentration before million/ml, Sperm motility %, morphologically normal sperm %, were Mean ±SD (3.10 ±1.23, 7.39 ±0.03, 46.67 ±15.31, 1.33 ±3.52, 36.00 ±5.07) respectively. Certain sperm parameters were highly improved after in vitro activation by PAF medium of Normozoospermic samples.
Background: Citrullus colocynthis (CC) is an herbal medicine used in the treatment of a wide range of diseases. Recently, It has been studied on in vitro activation of mice sperms with no research concern the effect of this plant on the DNA normality following in vitro preparation of human semen samples. Objectives: The present study was aimed to examine the effect of adding CC in the medium used for in vitro sperm activation of asthenospermic men and related factors and to found out the effect of CC on the DNA status following the activation. Materials and Methods: Citrullus colocynthis aqueous extract (0.01mg/ml culture medium) was used for in vitro sperm activation . One hundred and ten semen samples were divided into two groups, the first group activated in vitro by adding CC extract to the culture medium Ham's F-12 (the treated group) and, the second group sperm activated in vitro with Ham's F-12 alone ( the control group). Results: In vitro activation of sperms with aqueous extract of CC has shown positive effect on sperm concentration, sperm motility, and grade activity of progressive forward movement. There was a significant (P<0.05) increase in certain sperm function parameters of asthenozoospermic in treated group compared with the control group .The study showed that the DNA fragmentation resulted from treated group were lower than that of control group. Conclusions: According to the results of present study, adding aqueous extract of CC to the human sperms activation culture medium leads to an improvement in certain sperm function parameters of asthenozoospermic men with no harmful effect on the human sperms DNA with decrease in DNA denaturation.
Cigarette smoke contains a substance which is a toxic mutagenic chemical, and carcinogenic compounds, which have a negative effect on male fertility. Smoking cigarettes is associated with noticeably higher levels of the oxidative stress-causing reactive oxygen species in the seminal fluid. Because of this, smoking may cause changes to the sperm plasma membrane and a significant amount of deoxyribonucleic acid fragmentation. To study the influence of smoking on sperm chromatin immaturity and ICSI outcomes (fertilization rate, embryo grading and pregnancy rate). At the laboratories of the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies and after 1-7 days of sexual abstinence, 92 males’ sperm samples were taken. After liquefaction, macro-and microscopic studies were carried out, criteria applied by the WHO 2021. Masturbation was used to obtain the sample, which was then placed in sterile containers. The relationship between the smoking and chromatin immaturity and ICSI outcomes have been evaluated. A significant positive correlation between smoking status and sperm chromatin immaturity. Furthermore, there was a negative non-significant correlation between smoking status and fertilization rate. Weak and non-significant correlations between smoking status and embryo grade 2 as well as embryo grade 3. There was a positive association between smoking status and embryo grade 1. The sperm chromatin integrity was significantly lower in the non-smoker group compared to the smoker group.
Background: Male obesity is associated with physiological changes that can negatively affect male fertility. Obese men have demonstrated a 3-fold increase in the prevalence of oligozoospermia when compared with men with normal Body Mass Index (BMI). Objective: To estimate the role of aromatase enzyme in decreasing sperm parameters in obese infertile men. Subjects, Materials and Methods: Seventy obese infertile male (patients) and twenty obese fertile male (control) are enrolled in this study. Seminal fluid analysis done according to criteria of WHO (2010). Results: A significant negative correlation was found between human cytochrome enzyme and sperm concentration, progressively motile sperm, non-progressively motile sperm, immotile sperm and normal morphology. Conclusion: The present study showed clearly that male obesity negatively impacts fertility through direct association between increased (BMI) and semen parameters. The results suggest that increased estrogen as a result of aromatization in the fatty tissue, reflected through the increase in human cytochrome enzyme (aromatase), may be an important mechanism for the hypoandrogenemia and altered sperm parameters.
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.
Background : inflammasome protein govern the inflammatory process. The ele vation of NLRP3 inflammasome might be related to bad seminal fluid analysis results. Objectives: This study aimed to evaluate seminal plasma NLRP3 protein and its relation with seminal fluid analysis parameters before and after in vitro sperm activation. Patients and Methods: a total of 50 infertile patients were collected form higher institute of infertility diagnosis and assisted reproductive technique, there se men samples were processed for seminal fluid analysis before and after in vitro sperm activation. NLRP3 inflammasome protein were measured in seminal plas ma. Results: the mean level of NLRP3 protein was 5.12 ng/ml. the higher level of NLRP3 protein (4.5ng/ml) associated with lower sperm concentration, grade A, B, C and higher level of grade D motility. However, 50% of those patients have an improvement in their seminal fluid analysis after in vitro sperm activation. Conclusion: Elevation of NLRP3 protein in seminal plasma may be associated with the increased leukocytospermia, bad sperm quality and unresponsiveness to in vitro sperm activation among infertile male.
Background: Both clomiphene citrate and tamoxiphene citrate, as an anti estrogen, is the most common prescribed medication for male infertility.In humans, ant estrogens interfere with the normal negative feedback of sex steroids at the hypothalamus and pituitary, thereby increasing endogenous gonadotropin releasing hormone (GnRH) secretion from the hypothalamus and follicle stimulating hormone (FSH) and luteinizing hormone (LH) secretion directly from the pituitary. Objective: The objective of this study was to investigate the effect of clomiphene citrate (CC) and tamoxiphene citrate (TC) administered to male mice on quality of sperm motility and activity. Materials and Methods: Ninety six male mice were divided into two major groups, and every one of these groups was subdivided into three minor groups involving control and two treated groups. In the CC study, 48 mice were administrated orally 0, 0.0125 mg and 0.025 mg as control (G 1), low dose (G 2) and high dose (G 3) for 42 days. While, in the TC study, 48 mice were administrated orally 0, 0.01 mg and 0.02 mg as control (G 1), low dose (G 2) and high dose (G 3) for 42 days. Then, the male mice were sacrificed, both vas deferens were collected and assessment sperm motility and activity. Results: The results of present study appeared that the use high dose of CC or TC significantly increment (P<0.05) sperm motility and activity as compared to control groups. Conclusions: Based on the results of this study, it can be concluded that the administration of high dose of either CC or TC increases sperm motility and activity.
Background: The semen cryopreservation has an important role in the male fertility preservation. Certainly, patients who are prone to become infertile due to surgical or medical treatments such as chemo- or radiotherapy for cancer treatment. In fact, semen cryostorage seems to be the only proven method that may offer these couples a chance of having children in the future. Objectives: This study was aimed to investigate the effects of cryopreservation protocols on human sperm parameters. Methods: Sixty semen samples with range of age (19-35) years were included in this study. Sperm parameters, were assessed pre- or post-cryopreservation. Cryopreservation protocols were done using SMART medium with either 15% dimethyl sulfoxide(DMSO) or 15% glycerol alone (as control group) or supplemented with either 0.25M or 0.5M of sucrose for treated groups. Crude data were statistically analyzed. Results: Generally, the results of the present study showed that the sperm parameters were significantly deteriorated post-thawing (P<0.05) as compared to precryopreservation. Both treated groups (G2 and G3) showed best results as compared to G1 (control groups). In contrast, treated group (G2) with 0.25 M of sucrose appeared better results than the as G3 group that treated with 0.5M of sucrose. Conclusion: Sperm parameters including concentration, motility, and morphology were deteriorated after cryopreservation. Using 0.25M sucrose reduced the impact of cryopreservation.
In about 40% of infertile couples, the husband was found to be either the single or a contributing cause of infertility and the systemic management ready for those couples with male infertility are restricted. 135 couples with infertility shared herein; the husbands required in vitro activation for their sperm samples, their wives however; had no fertility problem and were submitted ovarian induction protocol. However; Sperm preparation was done using both, the centrifugation indirect swim-up technique with Global medium and density gradient with ALLGrad solution. While ALLGrad 45% is a ready-to-be-used upper layer. There was an acceleration in seminal parameters after preparation using both various techniques. So, a highly significant increment (P≤0.001) in the percentages of progressive forward motility was in grades (A and B) and grade (A) progressive sperm motility, also a significant enhancement (P≤0.05) in percentage of morphologically normal sperms when applying density gradient mechanism in relation to centrifugation swim-up technique. This study concludes that the use of density gradient mechanism with ALLGrad solution and Global culture medium are more efficient in achieving better percentages of progressive forward motility grades (A and B), grade (A) progressive motility and morphologically normal values of spermatozoa than centrifugation indirect swim-up technique by using Global culture medium. However, it was found there was a non-significant difference when we use both techniques concerning other sperm parameters.
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.
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.
The ability of human sperm to fertilize can be compromised both in vivo and in vitro by the toxins found in cigarette smoke, which can lower sperm mitochondrial activity and harm chromatin structure. The aim is to study the effect of smoking on the apoptotic marker (caspase-3) of human sperm. A cross-sectional study of 80 semen samples of sub-fertile men, were divided into smokers (35 patients) and non-smokers (45 patients). All the semen samples after seminal fluid analysis underwent HOST test and grading (A-G) according to the WHO 2021 criteria then the slides were smeared and stained with an immunocytochemistry kit for Caspase-3 apoptotic marker. There was no significant effect of smoking in the concentration of caspase-3 expression, in spite of the high presentation of both grades A and G in the smoker group ( p = 0.57, 0.56 respectively). Smoking has a negative impact on the DNA integrity of sperm as it increases the apoptotic marker within the smoker group.
Background: Many studies conducted on the pentoxifylline (PX) and Glycyrrhiza glabra (G.glabra) as motility stimulants showed positive effect on sperm preparation in vitro improving the progressive forward movement. However, it was not known the impact of these stimulants on the nature of genetic material, especially after cryopreservation. Objective: The experiment was designed to find out any harmful effect of a medium containing a mixture of PX and aqueous extract of G.glabra on the sperm DNA before and after cryopreservation. Materials and Methods: Fifty mature fertile male mice 8-12 weeks old were used. Epididymal sperms were obtained from caudal region and prepared in vitro by direct activation technique using four media namely; PX alone, G.glabra alone, a mixture of PX with G.glabra and Ham's F-12 (as control medium). Certain sperm function characteristics were examined as well as evaluation of DNA integrity by acridine orange test and comet assay before and after cryopreservation. Results: The results showed a highly significant (P<0.001) improvement in the sperm forward movement, with a highly significant (P<0.001) decrease in DNA abnormality using acridine orange test and comet assay, following in vitro activation by the four media compared to before activation. Following in vitro activation and cryopreservation, the results of epididymal sperm showed a highly significant (P<0.001) decrease in DNA abnormality by using PX medium alone, G.glabra medium alone and the mixture of PX and G.glabra compared to control medium and the results before cryopreservation. Conclusion: According to the results of present study, it has been found no toxic effect of medium containing PX and/or G.glabra on the epididymal sperms DNA after activation in vitro and cryopreservation using acridine orange test and comet assay. The best result obtained when using a medium containing a mixture of PX+ G.glabra together. These data could be applied for preparation and activation of epididymal and testicular sperms obtained from obstructive azoospermic men.
Background : At least in 25% of infertile men, no identifiable cause can be at tributed to the problem,so it is termed idiopathic infertility. Those men are candi dates for empiric medical therapy which include antioxidant therapy. Men upple ment Plus (MFS Plus) uses a powerful all-natural formula of many antioxidants in addition to Relora to improve semen quality. Objectives: To evaluate the effect of MFS Plus on sperms concentration, motility and morphology in men with oligo and/or asthenozoopermia in sample of Iraqi infertile males. Methods: Acomparative clinical trial study was conducted at High Institute of Infertility Diagnosis and Assisted Reproductive Technologies during the period from July 2015 till February 2017. Male partner of infertile couples who had one or more of the following criteria oligo, astheno, and/or teratozospermia accord ing to criteria of WHO 2010 were involved in this study. Eighty-one males were divided randomly into two groups, the first group (group A) were given MFS Plus capsule AMS® (America Medic & Science) two cap sules twice daily for three months and the second group (group B) were given vitamin E capsule (one capsule three times daily) for the same period.A second semen analysis were done after three months for each subject in both groups to show the effect of both types of treatment on semen quality. Results: There was a highly significant increase in sperms progressive motility % (14.0±11.41 vs 29.35±15.61, p value < 0.001) and a highly significant decrease in immotile sperms % (60.15±19.3 vs 41.52±18.18, p value < 0.001) in the group of patients who received MFS Plus treatment. Moreover, there was significant increase in sperm concentration and percentage of normal sperm morphology (p value = 0.027, 0.019 respectively). However, no significant change was observed regarding semen volume and percentage of non-progressive motility.Whereas there were no significant changes in any of semen analysis parameters in the group of patients (group B) who received placebo. Conclusion: MFS Plus usage for three months is very beneficial for sperm motil ity in addition to its valuable effect on sperm concentration and morphology and it is more efficient than vitamin E alone.
Background: Cadmium (Cd) is a heavy metal toxicant, present widely in our environment and workplaces.It is well known that cadmium causes adverse effects on male reproductive organs and reproductive functions in experimental animals. Objective: This experiment was designed to find out the effect of cadmium on some histological and hormonal parameters of the reproductive system of adult male rats. Methods: thirty mature albino male rat were used (Rattus rattus norvegicus albinus), as a mammalian model. these animals were divided into two groups, 15 rat / group after labeling them. The control group (G1) was orally administrated with distilled water during the 4 weeks period of the experiment and the experimental group (G2) was orally administrated with 20 ml cadmium chloride (200 mg/1L body weight) during the 4 weeks. First of all, the rat were anesthetized, after that blood aspiration using anesthetic cardiac puncture and the serum was separated and store at -20°C until assessment of FSH, LH and testosterone hormones. After that, the weight of reproductive organs (testis,epididymus and seminal vesicle) was recorded. These organs were fixed with 10% formal saline for histological study. Results: The statistical analysis shows a significant (P< 0.05) decrease in the weight of the testes and seminal vesicles when compared with control group. The results of the sperm parameters show a significant (P< 0.05) decrease in sperm concentration and a highly significant (p≤0.01) decrease in sperm motility in treated group as compared to control. The result of testicular sections in treated group shows a decrease in thickness of germ cells layer, widening of the central seminiferous tubules lumen and prominent germ cells population necrosis. Multiple vacuoles were seen within the tubules. Sertoli cells were abnormal in number and shape as compared to control. A peritubular fibrotic change had been seen also in the testicular sections. The result also shows a highly significant decrease in diameters of seminiferous tubules, primary spermatocytes, secondary spermatocytes, spermatids and increase in interstitial space when treated with cadmium compared with control group. The Levels of serum hormones shows a significant (P<0.05) decrease in the levels of testosterone and LH hormones mean in the treated group,while highly significant (ps0.01) increase in the FSH hormone mean in the treated group when compared to the levels mean of the hormone in the control group.. Conclusion: In conclusion, these findings suggest that cadmium administration with 20 ml cadmium chloride (200 mg/1L body weight) for 4 weeks during the adulthood induced damage in the reproductive organs and lead to impair the functions of sperm parameters and cause histological and hormonal alterations.
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 %.
Swim up is the most popular technique of sperm preparation, in which sperm is selected on motility and ability to swim out the plasma of the semen. The goal of this research is to compare the influence of adding platelet rich plasma (PRP) and phosphate buffer saline in semen prepared by swim – up technique. This research included 119 semen sample evaluated according to sperms parameters stated in WHO and divided into 2 groups. The first group treated with equal quantity of phosphate buffer saline (250) μm contain 10% human serum albumin (HSA) while second group treated with equal quantity of phosphate buffer saline (250) μm contain 10% human serum albumin (HSA) and 2% platelet rich plasma. The samples are prepared by swim – up method. Applying swim up technique using-PBS+PRP (HAS) medium resulted in highest sperm parameters in a highly significant manner (p < 0.001). The values were far better in Swim -up PBS (HAS) with added 2% PRP than those produced by swim up –PBS (HAS) alone. The statistical analysis shows that the variations between Swim up-PBS (HAS) with addition 2% PRP media and swim -up PBS (HAS) were highly significant.
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: Tannery work share exposure to solvents and chemicals , which has been linked with inverse pregnancy and fertility outcomes in human being as reported in some studies . Objective: The current study aimed to through some light on the harmful pollutant chemicals effect on tannery workers reproductive health. Literature in this portion are rare in Iraq, so it had been tested the hypotheses that leather workers suffered from male infertility parameters due to the continuous exposure to chemicals that used in this industry. Subjects, Materials and Methods: This study included 60 subjects can divided into 2 groups: 30 males as study group and 30 males as control group, the period of study was between October 2014 to April 2016, the samples were taken from tanneries workers from various governorates of Iraq. Results: The data clearly showed a significant infertility parameters among these workers represented by low total sperm number, low sperm activity the significance levels were within the value of P ≤0.05 accompanied with a significant high seminal fructose. Conclusion: The main conclusion was that chronic exposure to these chemicals in tannery workers especially Cr VI and N,N-dimethyle form amide may cause male infertility parameters.
Background: Oligozoospermia is one of male infertility type which related with sperm lowering count in seminal fluid under limited reference. There are many factors leads to oligozoospermia like, congenital defects, endocrinal disorders, genetic abnormalities, immunological factors , life style , or in some cases unknown reasons. Immune and some non-immune cells produce immuno-chemical messengers proteins called (Interleukins) work as alerts proteins and contribute in immune body defend. These mediators found in all body fluids, also found in seminal fluid,and it’s presence of certain concentrations reflects the state of health of the body. Objective:Measure the level of interleukin-6 in fertile (normospermia) and infertile (Oligozoospermia). Materials and Methods: Microscopically and macroscopically ejaculates analysis according to WHO (2010) criteria for eighty -seven ejaculate from normospermia and oligozoospermia patients with age rang (21-47)yrs. , in duration of ( Nov-2013 to Mar-2014), study divide in to three groups according to the concentration ranges in to (normospermia > 15X 106 cells/ml), (Mild to Moderate Oligozoospermia 15-5 X106cells/ml) , (Severe Oligozoospermia< 5X106). Then store all samples in(-20 ) , IL-6 was detected through the (ELISA) specific way and kit . Results: Interleukin-6level is less in normospermia group than Oligozoospermia groups at (P<0.05), IL-6 level not correlated with sperm count, except weak correlation in severe group at (P≤ 0.05) and (t α=0.05). Conclusions: There are no correlation between IL-6 level and sperm cells concentration but the variation of interleukin-6 level among the groups indicate that some sperms parameters influence on interleukin-6 levels among the groups like sperm motility or morphology and also that the causative factors which lead to oligozoospermia also effect on IL-6 level and effect on spermatogenesis .
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.
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.
For couples with a variety of male factor disorders, intracytoplasmic sperm injection has made pregnancy and motherhood possible. Two hundred-one males who had non-obstructive azoospermia or severe oligospermia were included in this study. One hundred forty trials of ICSI were done by using fresh sperms (fresh group n=140), while 61 trials of ICSI were done by using cryopreserved sperms (cryopreserved group n=61) to compare the conception rates between fresh and cryopreserved sperms used in intracytoplasmic sperm injection (ICSI). Twenty-five samples from each group were collected by doing a testicular biopsy. Pregnancy rates were higher using cryopreserved sperms as compared to fresh spermatozoa, 39.3% compared to 24.3% in the fresh group ( P .030), respectively. Binary logistic regression analysis showed that the odds ratio of achieving pregnancy with cryopreserved sperms was 2.022 higher than with fresh ( P .030). Cryopreservation has no negative effect on the pregnancy rate. Instead, it might improve the results of ICSI in comparison with a fresh sperm sample.
Receptive endometrium is essential for successful embryo implantation and is created by the synergistic activities of estrogen and progesterone, as well as support from other endocrine, paracrine, and autocrine pathways. Granulocyte Colony Stimulating Factor (G-CSF) may play a crucial role in modulating the immune response to enhance implantation. To estimate the role of granulocyte-colony stimulating factor on the clinical intracytoplasmic sperm injection (ICSI) outcome parameters. A prospective comparative study with random sample selection was carried out from 2021 to 2023 at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University. It included 80 women with infertility, with or without previous failure trials of intracytoplasmic sperm injection, who completed IVF and ICSI protocols and reached the day of embryo transfer. On the day of embryo transfer, patients were divided into two groups: 49 patients received no additional treatment (group 1), and 31 patients received granulocyte-colony stimulating factor subcutaneous injection within one hour of embryo transfer (group 2). The implantation rate was 13.3% in group 2 compared to 7.6% in group 1. The chemical and clinical pregnancy rates were 29.0% and 25.8% in group 2, while group 1 had 18.4% for both chemical and clinical pregnancy rates. The pregnancy and implantation rates were slightly higher in group 2, but the difference was not significant. More research is needed to support the use of G-CSF as a standard treatment in ICSI patients.
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.
Background: Human growth hormone (HGH) is undisputedly a potent hormone with a wide variety of biological effects. The anabolic actions of human growth hormone have made it attractive as a potential agent for catabolic problems in a wide range of clinical conditions. Objectives: In the present study, we investigated the possible relationship between levels of HGH in serum (S. HGH) and seminal plasma (Sp. HGH) and semen parameters in normozoospermic (NZS) men and infertile patients. Materials and Methods: In total, 66 infertile patients with azoospermia (AZS; no. 52), asthenozoospermia (AsZS; no. 6) and oligoasthenoteratozoospermia (OATZS; no. 8), and normozoospermic men (NZS; no. 12) were included in this study. From each subject, semen and blood samples were taken to assess seminal plasma and sperm parameters and levels of S. HGH and Sp. HGH; respectively. Results: The highest S. HGH was evaluated in AZS patients when compared to NZS and other infertile groups. Significant elevation (P<0.05) in the level of S. HGH was assessed for AZS patients as compared to NZS men. In contrast, the level of Sp. HGH in NZS men was significantly elevated (P<0.05) when compared to AZS patients only. Positive and significant correlations were noticed between levels of Sp. HGH and semen liquefaction time in NZS, AZS and OATZS groups. In NZS men, positive and significant correlation were observed between Sp. HGH and semen volume and negative and significant correlations were detected between Sp. HGH and percentages of sperm motility, progressive motility, immobility and agglutination. Conclusion: Our study represents the first study in Iraq showing that the level of HGH in serum and seminal plasma affect male fertility and sperm parameters in normozoospermic males and infertile patients. Further biochemical and molecular studies are recommended to investigate the mode of action of HGH and its receptors in relation to fertility.
Non-obstructive azoospermia (NOA) is the absence of sperm in infertile men ejaculate as a result of significant spermatogenesis defects or hormonal malfunction. PRP intratesticular injection may be employed in assisted reproductive technology as a therapeutic effect for this problem. The aim is to assess the level of spermatogenesis and reproductive hormones after PRP autologous intratesticular injection. 50 non-abstructive azoospermia infertile men participated in the current study. The patients were given an intra-testicular injection of platelet-rich plasma using a 1 ml syringe; FSH LH hormonal levels were estimated; if there was no sperm count, the patient should be prepared for a testicular biopsy by the expert doctor. Small number of sperm may be discovered during a biopsy while the testes are being examined in the lab. The therapeutic benefits were seen in 15 patients 3-4 months after using autologous platelet-rich plasma. The results showed that there was a highly significant difference in FSH and LH hormonal levels after PRP injection P(0.001). PRP can help non-obstructive azoospermic infertile males with their testis' structural and functional dysfunction.
The toxic effect of drugs and environmental chemicals on the human reproductive system has become a major health concern. Lead is recognized as the most dangerous of the toxic metals and a major threat to the biological systems of both human and animals. In this study we detect the effect of two types of antioxidants: flaxseed and zinc, on the toxicity of low dose of lead acetate on reproductive organs of male mice, this study was conduct. Two groups of mature male mice,exposed to low dose of lead, were administered either zinc sulphate (0. 5mg/kg body weight/day) or flaxseed oil (2.5% of total daily food intake), for 6 weeks. The results showed that lead caused a significant reduction in the total body weight, testicular weights & adverse effects on sperm parameters, while the concomitant administration of antioxidants caused elevation in these weights with significant improvement in sperm quality as compared to the only lead treated group. It is concluded that lead, like all other heavy metals, could induce oxidative stress and the concomitant administration of antioxidants (zinc or flaxseed) can ameliorate the toxic effect of lead on reproduction.
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.
In human-assisted reproduction both partners need to be evaluated during counseling because causes of infertility are due to male factors and female factors. some body hormones play important role in fertile women and cortisol one of these hormones which is essential for normal ovulation, cortisol usually increases during the follicular phase of the menstrual cycle, play a role in the early stages of follicular development by their interaction with aromatase activity and granulose cell functions. To investigate the correlation of cortisol hormonal level in serum and in follicular fluid that relationship with age in women under Intracytoplasmic sperm injection outcome. Its randomized prospective clinical trial study performance conducting in the Higher Institute for Infertility Diagnosis and Assisted Reproductive Technologies / Al-Nahrain University, Baghdad-Iraq, during the period from November2021to June 2022. a total of Sixty-three Infertile Iraqi women, who planned to inter intrasytoplasmic sperm injection program. The blood sample and follicular fluid sample obtained from all infertile women inter intracytoplasmic sperm injection program to measure cortisol hormonal level at specific days of this study. There was increase in cortisol level in women aged between (30-40) years, So there was significant finding between cortisol level and this age group and there was increase in cortisol level in women aged < 30 years at the same day and no significant difference in cortisol level in women age more than 40. Significant differences in cortisol level appear in group of women age between (30-40) years, also the same result appears in group of women age more than > 40 years and no significant differences in cortisol level in women aged < 30. No significant differences in cortisol level in all age women group (30-40) years, <30 years ,and the women age > 40 years.
The structural elements like the “blood-testis barrier” (BTB) and systemic and local tolerance mechanisms help to maintain testicular homeostasis, which shields germ cells and mature sperms from immune attack. Any damage to this barrier could result in an immune attack and may lead to infertility. This study aims to look at the correlation between male infertility and immunological factors (IL-17, IL-25, and anti-sperm antibody). During the months of October 2021 and July 2022, a total of 144 samples were taken from patients who sought to have infertility tested. The subjects' semen specimens were taken in order to perform the ELISA tests for IL-17, IL-25, and ASA. The levels of ASA, IL-17, and IL-25 differed significantly (P≤ 0.05) across study groups, according to the results of immunological parameters. The results of the correlation analysis revealed a strong positive association between IL-17 and IL-25, but not between those two interleukins and ASA. Although there was theoretically a positive link in this study, it was very weak and not significant. The findings suggest that IL-17, IL-25, and ASA influence seminal fluid parameters, which in turn affect male fertility.
Background: The intra uterine insemination (IUI) still an easy, simple and effective method to overcome the infertility that resulted from different causes. Although the equipments and methods of assisted reproductive technologies (ART) were highly developed, the percentage of successful IUI live-birth not exceed 20-30%. Objective: The present study was aimed to elucidated the cut off values of in vitro sperm preparation technique, follicular number and size, endometrial thickness and hormonal profile that lead to successful pregnancy rate (PR)following IUI. Materials and Methods: In retrospective study, one hundred cases of infertile couples were in who were became pregnant following IUI at the Institute of Infertility Diagnosis and Assisted Reproductive Technologies, through the period from January 2007 to January 2010. Depending on the results of retrospective study, IUI was achieved for 100 infertile patients between August 2010and June 2011. The mean of significant prognostic variables were measured in both studies namely: age, type of infertility, type of sperm activation techniques, the protocol of ovulation induction medicine, the mean of hormonal status of Luteinizing Hormone (LH), Estradiol(E2), and number with diameter of follicles, endometrial thickness a cycle day before hCG injection. Results: The protocol of ovulation induction in IUI cycles by using clomiphene citrate(CC) and recombinant gonadotropin (rFSH) was significantly(p<0.05) improve the IUI results compared to CC alone or CC with PergonalS. The mean values obtained by ultrasonography namely; two follicles, and dominant follicle of 19.5mm in size with endometrium thickness of >9.5mm were gave significantly successful IUI outcome compared to other values. In prospective study similar results were obtained, therefore the overall pregnancy rate was 37% per cycle. Conclusion: The calculated data of successful IUI outcome from retrospective study gave a best pregnancy rate in prospective study which was 37%. Consequently, these results can be considered as guideline for physician who interest in assisted reproduction to have a success in IUI.
Background: Carnitines are widely distributed in nature and their potential health benefits have been popularized. The carnitines exert a substantial antioxidant action. It is often sold as a nutritional supplement. The use of carnitine showed some promise in treatment of male infertility. Objectives: This study was designed to investigate the effects of L-carnitine (LC) administered to pregnant mice on some reproductive parameters of male offspring. Methods: Ninety mature male offspring mice (age: 6 weeks) were divided originally into two groups depending on the pregnant mothers were administered either 0.5 mg/Kg L-carnitine (Treated group) or distilled water (Control group) throughout pregnancy period. Whole body and testicular weight, diameter of seminiferous tubules, thickness of spermatogenic cell layers, levels of serum gonadotropins and sperm parameters were assessed. Results: In the present study, weight of whole body and testes of the male mice offspring group where mothers treated with 0.5 mg/Kg L-carnitine throughout gestation period was significantly increased (P<0.05) as compared to the control group. Further, significant increment (P<0.05) was observed in the thickness of spermatogenic cell layers and diameter of seminiferous tubules for the treated group as compared to the control group. Also, levels of serum follicle stimulating hormone (FSH) and Luteinizing hormone (LH) in the treated group showed significant elevation (P<0.05) as compared to the control group. Sperm parameters for the treated group were enhanced significantly (P<0.05) as compared to the control group. Conclusion: Administration of low dose of L-carnitine to pregnant mice improved reproductive performance of male offspring. Further molecular studies are recommended to investigate the effects of L-carnitine administered to pregnant mice on quality and quantity of acrosomal enzymes for male offspring.
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.
Various types of anesthetic techniques were described for oocyte recovery. Different anesthetic agents have been used with different effects on oocyte quality and embryonic development and studies on the potential toxicity of general anesthesia ended up with conflicting results. Many experiments as well as human studies documented the existence of anesthetic agents in the follicular fluid (FF) with possible adverse effects on the oocyte quality and subsequent embryo development. Ketamine and remifentanil were used for the studied groups to compare oocytes and embryos characteristic of infertile women during oocytes retrieval under general anesthesia: one group receiving ketamine, midazolam, and propofol and the other group receiving remifentanil, midazolam and propofol. 60 infertile females were undergoing intracytoplasmic sperm injection categorized in two groups, group I received midazolam, remifentanil and propofol; whereas, group II received midazolam, ketamine and propofol. There was highly significant difference in mean abnormal oocyte between ketamine and remifentanil groups. Grade III embryos were significantly limited to ketamine group. FF ketamine and remifentanil were not significantly correlated to any of oocyte characteristics or embryo characteristics. Anesthetic agents, ketamine and remifentanil, were detected in the FF of infertile women and within recommended doses. Ketamine resulted in adverse effects on oocyte and embryo quality in comparison with remifentanil represented by greater percentage of abnormal oocytes and that bad quality embryos were limited to ketamine group.
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: Azoospermia is defined as the lack of sperm in male ejaculate and is suggested to be present in about 1 % of all males and to account for 10-15% of the causes of infertility in men. The development of intracytoplasmic sperm injection (ICSI) as an effective therapy. Embryo grading has gained special attention in published literature regarding its predictive role in pregnancy outcome following ICSI; however, substantial controversy is present that justified the carrying out of the present study. Objective: To study the association between embryo quality and pregnancy rate in women of males with azoospermia. Patients,Materials and Methods: This prospective cohort study included 60 patients with azoospermia, 15 male with obstructive type and 45 males with non-obstructive type.Those patients were randomly selected form the population of infertile couples who regularly visits the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies and from Kamal Al-Samarrai Hospital -Center for Infertility Treatment and IVF. Results Pregnancy was encountered in 7 patients (23.33%). It was not statistically associated with type of azoospermia (P=0.388), also it was not affected by age and serum hormonal level, but highly determined by good quality of embryo. Quality of embryo was graded from 1 to 4. The lower the grade, the better the quality of the embryo was. Quality of embryo was not significantly correlated with type of azoospermia, age and hormonal levels. Conclusions Good quality of embryo is a significant predictor of positive pregnancy outcome in ICSI and the quality of embryo is not affected by age of male patient or type of azoospermia.
Background: Infertility is a common social and health problem. Male factor is a major cause in 30% of infertile couples. Smoking is common at reproductive age and its effect on fertilitiy is still in controversy. Studying its effect on fertility helps in the management of infertility. Objectives: To assess the effect of smoking on serum sex steroidal hormones as well as on seminal plasma (SP) IL-2, IL-6 and conventional semen parameters in infertile men and their correlations between each other. Methods: Seventy male partners of infertile couples (30 smokers and 40 non-smokers) were involved in this study. The man was considered smoker if he had history of smoking of more than 10 cigarettes per day for at least one year. Semen analysis was done according to world health organization (WHO) 2010. Specific kits were used for measurement of serum testosterone, serum estradiol II, SP IL-2, and SP IL-6. Results: The median of SP IL-2 was significantly (p<0.05) higher in smokers than in non-smokers group and positively correlated with SP IL-6 in them. In addition, IL-2 was negatively correlated with semen volume, sperm motility and morphology, while IL-6 was negatively correlated only with sperm motility. However, there was no significant difference of serum sex hormones level or on semen parameters between smokers and non-smokers. Conclusion: Cigarette smoking affect cytokine levels in the seminal plasma but has no effect on serum sex hormones level or semen parameters.
Background: Infertility is considered one of the main public health issues, as it affects about 15% of the couples of reproductive age. The male factor is involved in 40% - 50% of infertility cases. It is difficult to assess accurately the overall magnitude of the contribution of genetics to infertility as most, if not all, conditions are likely to have a genetic component. The genetic causes of infertility are varied and include chromosomal abnormalities, single gene disorders and phenotypes with multifactorial inheritance. Some genetic factors influence males specifically, whereas others affect both males and females. Frequently, however, in the clinic no clear cause for the observed infertility could be diagnosed which at least in part, it reflects our still poor understanding of the basic mechanisms that regulating and controlling the genetic networks of male infertility. Objective: To study the molecular identification of Azoospermic infertile patients by the gene sequencing of NR5A1. Subjects, Materials and Methods: A study carried out during the period from November 2014 to June 2015. Fifty specimens were collected from thirty azoospermic patients and twenty normal healthy subjects (normozoospermic subjects) as control their age ranged between 23 and 48 years old, the seminal fluid of cases indicated that liquefaction time, color and viscosity equal in normozoospermic subjects (control group) and patient group while there is significant difference P≥0.05 in sperm count reach to more than 20 million and motility reach to more than > 25% in control subjects while there is no sperm count found in azoospermic patients. Then blood was collected for hormonal assay, in azoospermic patients the results revealed a gradual decrease in serum testosterone levels with a concomitant increase in serum follicular stimulating hormone (FSH) level when compared with control subjects . NR5A1 gene were investigated in 30 samples of extracted DNA from azoospermic patients by using polymerase chain reaction (PCR) method it can directly detect the NR5A1 gene content after it had been molecularly identified then were sent successfully for sequencing analysis ,this study examined the presence of two transversion and one transition mutation of NR5A1 gene in azoospermic patients compared with control subjects, there’s no mutation in the same gene when compared with gene bank.
Granulosa cells produce most ovarian function indicators in infertile women like estradiol, AMH, and inhibin B. Female peptide hormone insulin-like factor 3 is produced by theca cell. Signaling of insulin-like factor 3 from theca cell stimulates antral follicle development and theca androgen production. To investigate how INSL3 levels may be a theca cell marker of ovarian reserve and the association with current markers of ovarian reserve, including AFC, FSH, E2, and AMH, in infertile women undertaking assisted reproduction. In this cross-sectional study, 75 Iraqi infertile couples were chosen at Al-Nahrain University's High Institute for Infertility Diagnosis and Assisted Reproductive Technologies from October 2022 to April 2023 for the ICSI cycle. Based on the levels of INSL3 in the serum on the day 2 of the menstrual cycle and on the day of retrieving oocyte in the follicular fluid. A comparison was made with current ovarian reserve markers (AFC, FSH, E2, and AMH) .From 75 infertile women undergoing intracytoplasmic sperm injection (ICSI), 21 of them achieved pregnancy. Positive Significant relations were found between serum INSL3 levels and anti-mullerian hormone (AMH and antral follicle count. there were no significant relationships between follicular fluids INSL3 with serum LH, FSH and E2 . The study provides compelling evidence for the significant role of INSL3 in female fertility and its association with the success of intracytoplasmic sperm injection (ICSI). Positive correlations between serum INSL3 levels and anti-mullerian hormone (AMH) and Antral follicle count AFC indicate its potential involvement in ovarian reserve and hormonal regulation.
Background : Bacterial vaginosis (BV)is a polymicrobial clinical syndrome re sulting from replacement of the normal hydrogen peroxide producing Lactoba cillus sp. BV is the most common lower genital tract disorder among women of reproductive age (pregnant and non-pregnant) and the most prevalent cause of vaginal discharge and malodor. Objectives: To study the role of asymptomatic bacterial vaginosis on outcome in patients undergoing ICSI. Patients and Methods: This prospective study was undertaken in the High Insti tute of Infertility Diagnosis and Assisted Reproductive Techniques / Al-Nahrain University. During the period from September 2017 – April 2018. A total of 100 infertile women .All of them were underwent controlled ovarian hyper stimula tion for intra cytoplasmic sperm injection cycle. Antagonist oocyte maturation used as ovulation induction protocol in all the cases. Assessment of oocyte mat uration and embryo quality is done to the all the cases, bacterial vaginosis smear was collected on the day of oocyte retrieval. B.V. smear diagnosis by Amsel’s Criteria and Nugent’s Scoring. Results: TThere was no significant difference in mean age of pregnant and non-pregnant ladies, 29.40 ±6.47 years versus 30.78±6.84 years, respectively (P= 0.333).In addition the frequency of women under 35 years age was compara ble in the two groups with no statistical significant difference, 34 (79.1%) versus 33 (73.3%), respectively (P = 0.528). Wide variation and non-normal distribu tion was the role followed by the duration of infertility concerning sub-fertile women participating in this study. However, no significant difference was ob served in the duration of infertility between pregnant and non-pregnant ladies, 6.00 (5.00) years versus 6.00 (5.00) years (P = 0.289). Comparison of basal hor monal levels between pregnant and non-pregnant groups revealed no significant difference despite the presence of some minor differences in mean hormonal levels between the two groups (P> 0.05). Comparison of oocytes characteristics between pregnant and non-pregnant ladies was conducted and revealed no signif icant difference in mean total number retrieved oocyte (TNO), ruptured oocytes (RO), abnormal oocyte (AO), germinal vesicle oocyte (GV), metaphase I oocyte and metaphase II oocytes (P > 0.05), table Mean total number retrieved oocyte (TNO), ruptured oocytes (RO), abnormal oocyte (AO), germinal vesicle oocyte (GV), metaphase I oocyte and metaphase II oocytes was 10.81 ±4.95 versus
The selection of protocols in controlled ovarian stimulation during intracytoplasmic sperm injection affect the treatment outcomes. Both GnRH agonist and GnRH antagonist protocols are clinically utilized, therefore comparision of these protocols is important for the evaluation of their laboratory and clinical outcomes.To compares the short GnRH agonist and GnRH antagonist protocols effectiveness in expected normal responder females undergoing intracytoplasmic sperm injection (ICSI). Methods: A retrospective study was involved 60 women aged less than 35 with antimullerian hormone (AMH) more than 2.5 ng/ml. Participants women were assigned to either group1 short GnRH agonist protocol and group2 GnRH antagonist protocol. The outcomes measured including total oocyte, mature and immature oocytes, number and quality of embryo and pregnancy rate.The short GnRH agonist protocol group showed statistical significant benefit in oocyte yield (p- value 0.04), metaphase I (p- value 0.09), metaphase II (p- value 0.03), embryo number (p- value 0.04), embryo quality (p-value 0.01) and pregnancy rate in expected normal responder women compared to GnRHantagonist protocol. The short GnRHagonist protocol reveals favorable ICSI outcomes than antagonist protocol and may be better choice in expected normal responder women.
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.
Infertile couples undergoing Assisted Reproductive techniques are really receptive to any intervention intended to enhancement their fertility. Successful implantation of an embryo requires a receptive endometrium, a good quality embryo, and embryo endometrial synchronization in all species. Pharmacological agents such as aspirin and sildenafil have been included in previous clinical trials to improve endometrial vascularity and perifollicular blood flow but the results were inconsistent. To evaluate the effect of oral low dose aspirin or vaginal sildenafil citrate on endometrial thickness, retrieved oocyte, degree of maturation, embryo quality and pregnancy test. This comparative prospective study was conducted on the 60 infertile women who participated in intracytoplasmic sperm injection (ICSI) cycles at the infertility clinic associated with the "High Institute of Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University / Baghdad/ Iraq." December 2020 to June 2022. In the present study the count of total oocytes retrieved was significantly higher in aspirin treated group in comparison to sildenafil treated group 12.5 ±5.3 versus 9.37 ±5.5 respectively. No significant variation in mean endometrial thickness among study groups. Regarding biochemical pregnancy test, positive results were obtained in 12 (40.0 %), 10 (33.3 %) in aspirin treated group, sildenafil treated group respectively and the difference was not significant. Aspirin treatment resulted in higher number of total retrieved oocytes, aspirin and sildenafil did not improve endometrial thickness, but they resulted in improvement of sub endometrial vascularity; however, biochemical pregnancy test was not affected by either pharmacological agent.
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.
Exposure to radiation of various forms is one of the many environmental, health, and lifestyle variables that have been linked in recent years to an increase in the rate of male infertility. According to data from several recently published studies, mobile phone use may negatively affect sperm parameters, which may lower male fertility. Ninety-seven samples of semen from fertile and infertile individuals were sent to the "High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Al-Nahrain University in Baghdad, between November 2022 and May 2023. The samples were split into four groups: normozoospermia (n = 40), asthenozoospermia (n = 25), oligozoospermia (n = 16), and teratozoospermia (n = 16). Each group was further divided into two groups based on the amount of time spent using a mobile phone each day: less than 4 hours per day and equal to or more than 4 hours per day. A routine seminal fluid analysis was carried out to measure the sperm parameters and test the deoxyribonucleic acid fragmentation rate using an acridine orange stain. Samples exposed to electromagnetic waves of cell phones showed a significant decrease in DNA normality. deoxyribonucleic acid fragmentation index (p < 0.001) in normozoospermia patients exposed to more than 4 hours/day of non-ionizing electromagnetic waves. The results of deoxyribonucleic acid fragmentation index after treatment were shown a significant improvement compared to before the treatment in all fertile and infertile groups. Electromagnetic waves emitted from cell phones are associated with a decrease of deoxyribonucleic acid normality when exposure for <4hours/day. The study recommend that men of reproductive age avoid keeping their cell phones in the hip pocket and front pants pocket which may negatively affect spermatozoa deoxyribonucleic acid integrity and impair male fertility.
The 5,10-methylenetetrahydrofolate reductase (MTHFR) enzyme is a key methionine and folate metabolism enzyme that is found in oocytes and embryos in preimplantation stages and plays a vital role in female reproduction.To assess the impact of MTHFR genetic polymorphisms (C677T and A1298C) on follicular stimulating hormone (FSH), luteinizing hormone(LH), Estradiol (E2), progesterone and antimullerian hormone (AMH) in female receiving intracytoplasmic sperm injection (ICSI), 85 infertile women undergoing ICSI treatment at the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies in Baghdad, Iraq, were recruited. These patients had their MTHFR polymorphism genotypes examined, FSH, LH, E2, progesterone, and AMH levels were compared between groups. Furthermore, all demographic characteristics as well as ICSI outcomes were compared between pregnant and non-pregnant women. The research revealed no discernible variation between the means of FSH, LH, E2 and AMH with respect to the 219A>C(rs1801131) (A1298C) genotypes (p > 0.05). Progesterone level was significantly differed with high level found in CC followed by AA and AC. There was no statistically significant variation in hormonal features between CC, CT, and TT for A1298C, 219A>C(rs1801133). The demographic features (age, duration, type of infertility, cause of infertility and previous trial) were not notably different between women who were pregnant and those who were not. In women undergoing ICSI, the maternal MTHFR polymorphisms is not related to FSH, LH, E2 and AMH. MTHFR polymorphisms therefore don't offer useful information regarding the hormonal levels in our groups.
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.
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.
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
It is well known that oocyte quality is essential in the determination of the developmental potential of the fertilized oocytes. The oocyte developmental potential decreases in all species with increasing age. Use of serum and follicular fluid concentration of GDF9 as biomarkers of oocyte and embryo quality and their relation to pregnancy rate. Eighty-eight women were included in this study are selected from those undergoing intra-cytoplasmic sperm injection. Positive pregnancy was achieved by 14 women. The difference in mean serum and follicular GDF9 between pregnant and non-pregnant women was not significant. MI oocyte count was not significantly correlated to serum and follicular GDF9 (p > 0.05). MII oocyte count showed a non-significant correlation to serum and follicular GDF9 (p > 0.05). Grade 1 embryo count showed a non-significant correlation to serum and follicular GDF9. Grade 2 embryo count showed a non-significant correlation to serum and follicular GDF9. Also, grade 3 embryo count showed a non-significant correlation to serum and follicular GDF9. The current study revealed that serum and follicular GDF9 could not be used as indicators for oocyte maturity, embryo quality, or pregnancy rate.
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
Iraq is one of the countries that have suffered from decades of heavy conflict, industrial activity, and little environmental regulation. As a result, notable environmental degradation has occurred. Urban environments, in particular, suffer severely from air pollution, which may be affecting reproductive health outcomes .This study examined the environmental pollution rates and infertility rates in 5 large urban areas in Iraq from 2019- 2023 .A cross-sectional ecological study was conducted across five large Iraqi cities. Air pollution data were obtained from monitoring stations and included PM2.5, PM10, NO2, SO2, and O3 measurements. Infertility rates were obtained from hospital records and fertility clinics. Statistical analysis was conducted employing Pearson correlation coefficients and multiple regression models .Overall, we found strong positive relationships between air pollution and infertility rates in both men (r=0.73, p<0.001) and women (r=0.68, p<0.001) with respect to PM2.5 concentrations. NO2 levels were also associated with decreased sperm quality parameters (r=-0.61, p<0.01). In areas where PM2.5 concentrations were >50 μg/m3, the infertility rates were 34% higher than those in areas with lower PM2.5 concentrations. Higher air pollution levels in urban Iraqi cities show associations with reproductive health outcomes. Meaning, there is an urgent need to address environmental health and policy reform in Iraq.
Pregnancy-Associated Plasma Protein (PAPP-A) is a zinc metalloproteinase in the insulin growth factor system (IGFs) produced by the syncytiotrophoplast region of the placenta. It plays a critical function in the cleavage of IGFBP4. In the ovary IGFs, it regulates follicular and oocyte maturation, and steroidogenesis. While in polycystic ovarian syndrome (PCOS) Hyperinsulinemia and hyperandrogenemia it causes follicular environment changes and early ovulation resulting in lower oocyte and embryo quality in patients and this will decrease the success of pregnancy in women enrolled in the ICSI cycle. The present study aimed to assess the relationship of PAPP-A levels in serum and follicular fluid in women with PCOS and non-PCOS with oocyte and embryo quality in women undergoing ICSI cycle. 45 infertile Iraqi women were enrolled. Women with PCOS had to meet at least two of the three criteria set by the Rotterdam ESHRE/ASRMS criteria, the age of the included women ranged between 20-45 years. In non-PCOS patients, PAPP-A has higher level in serum and follicular fluid but without a statistically significant difference matching with PCOS group. In addition, there was no significant correlation between PAPP-A levels in serum and follicular fluid with oocytes and embryo characteristics. However, PAPP-A levels are higher in serum and follicular fluid in women with positive pregnancy but without significant differences. PAPP-A had no correlation with oocyte and embryo quality.
Intratesticular testosterone plays a crucial role in male reproductive function and overall health. Body Mass Index (BMI) and smoking are known lifestyle factors that can influence hormonal balances. Elevated BMI has been associated with alterations in testosterone levels, impacting male fertility and endocrine function. The aim was to examine the concurrent relationships between intratesticular testosterone, BMI, and smoking habits. Biopsies of the testicles were taken from 32 patients with non-obstructive azoospermia. Testicular sperm extraction (TESE) under spinal or general anesthesia was performed after a comprehensive history, physical examination, and investigations. After performing a minced biopsy and centrifuging the fluid, testicular fluid was extracted. The highest intratesticular testosterone levels were in normal patients and lowest levels in over weight patient; on the contrary the highest serum 17-hydroxyprogesterone levels were found in over weight patient and lowest levels in normal weight patients; There were insignificantly higher both intratesticular testosterone and 17-hydroxyprogesterone levels in non-smokers. In conclusion, the statistical analysis revealed that there is an insignificantly low correlation between intratesticular testosterone levels, BMI, and smoking.
Glycotoxins termed as advanced glycation endproducts (AGEs) are made endogenously and exogenously. Maillard reaction is a chemical non-enzymatic process, between sugars and a free amino acid group of nucleic acids, proteins, and lipids which happens exogenously when handled with high temperature for a prolonged period and in a dry environment, resulting in glycotoxins (AGEs) formations. This reaction is responsible for the taste, color, and smell of food. Glycotoxins derived from food are absorbed in the intestine and store in cells and tissues. Also, glycotoxins are derived exogenously from smoking. While endogenous glycotoxins are generated by normal body physiology. Obesity and insulin resistance may induce glycotoxins (AGEs) accumulation. The objective of this study was to evaluate the relationship between glycotoxins (AGEs) and obesity. 66 infertile females who were undergoing intracytoplasmic sperm injection ICSI. These females were classified into three groups according to their body mass index. On the day of ova pick up the collect serum and follicular fluid for subsequent measurement of glycotoxins (AGEs) by ELISA kits. there was a significant difference in the concentration of glycotoxins in follicular fluid and serum between the three ICSI groups of patients. As well as there was a significant difference in the concentration of glycotoxins (AGEs) in follicular fluid and serum in the same ICSI groups of patients. there is a significant positive relation between glycotoxins (AGEs) and increase BMI.
Background: Speramax® was found to play an important role in sperm function characters and males reproductive performance with no studies on its effects on the oocyte maturation and embryonic development in females. Objective: The goal of the present work was to examine the effect of Speramax ® on oocyte maturation ,ova quality , embryonic development and live birth using mice as a model for mammals . Materials and Methods: In this study, Speramax ® was administrated orally for 1, 2 and 4 weeks . One hundred and ten female mice were randomly divided into four groups , the first group was superovulated(SUO) while the second group was treated by Speramax® with SUO and the third group was spontaneously ovulated (SPO) and treated with Speramax ® and the forth group was spontaneously ovulated (SPO ) and served as control group .Another forty-eight female mice were used for the determination of the number of live births . Results: The results indicated that treatment with Speramax® showed a positive effect on oocytes maturation in vivo. There was a highly significance ( p˂ 0.0001) improvement in the number of mature oocytes following treatment with Speramax® in SPO and SUO mice compared with SPO and SUO mice not treated with Speramax® . The percentage of embryonic development after 24 and 48 hours of mating in treated groups with Speramax® was significantly ( p˂ 0.05) higher than SPO and SUO mice .The study showed that the quantity and quality of embryos obtained from the treated groups were superior to that of the untreated group . Conclusions: It was concluded that the Speramax® greatly improved oocyte maturation, early embryonic development and embryo grading quality embryos with an increase in the numbers of mice live births.
Human chorionic gonadotropin is a glycoprotein hormone produced by the placenta. It exists in several isoforms with long half-lives (hours) and angiogenic potential; it maintains the corpus luteum to produce progesterone. This study aimed to evaluate Human chorionic gonadotropin concentration in follicular fluid and its effect on oocyte maturity and pregnancy rate, and whether it can be used as a noninvasive biomarker for predicting oocyte competence and pregnancy rate during intracytoplasmic sperm injection cycles. Infertile women undergoing ICSI treatment were included in the study. An antagonist protocol was used for ovulation induction. Patients were divided into two groups: pregnant women (n=18) and non-pregnant women (n=22). Samples of follicular fluid were collected on the day of oocyte pick-up. Chemiluminescent Immunoassay (CLIA) equipment was used for the quantitative determination of total β-human chorionic gonadotropin in follicular fluid. The mean beta-human chorionic gonadotropin level showed a highly significant increase relative to the mean level of non-pregnant women (110.24 ±20.50) versus (56.195 ±29.45). Moreover, there was no significant relationship between the levels of beta-human chorionic gonadotropin follicular fluid and the total number of oocytes and oocyte quality in the two groups. The Beta Human chorionic gonadotropin concentration in the follicular fluid plays an important and influential role in the prediction of pregnancy rates. Despite this, follicular fluid hCG level showed no relationship with the total number and quality of oocytes in the pregnant and non-pregnant groups.
Infertility is described as a couple's failure to conceive for at least a year, using unprotected sex. Even though all anesthetic chemicals have been revealed in the follicular fluids, general anesthesia is still used in many IVF centers for patients who want to get pregnant. This study included 60 infertile women randomly divided into two groups under general anesthesia. 30 patients were given a Ketamine dose of 0.5 mg/Kg and the other 30 patients were received Remifentanil dose of 0.5 µg/Kg for induction of anesthesia. We collected serum for Reactive Oxygen Species (ROS) levels assessment for all patients before and after starting general anesthesia. Routine ICSI procedures was performed on all participants, including clinical evaluation (history, examination, and investigation), controlled ovarian and ovulation stimulation, oocyte retrieval under general anesthesia, follicular fluid collection for postoperative anesthesia medication concentration (Remifentanil and Ketamine), oocyte stripping, oocyte maturity assessment, intra-oocyte sperm injection into mature cells (MII), fertilization and division evaluation and embryo categorization, embryo selection and transfer, luteal phase support, and beta-hCG determination. ROS concentrations were compared between the Remifentanil and Ketamine groups. There was no significant difference in embryo features between Remifentanil and Ketamine, indicating that neither one is superior to the other in this regard. When Remifentanil or Ketamine was taken, there was no significant difference in ROS levels in serum or follicular fluid.
Background: Leptin is an adipocyte —secreted protein that participates in the regulation of energy homeostasis. It prevents the body from storing fat, controls hunger and cravings, regulates food intake and energy expenditure, provides the body with an index of nutritional status and controls the whole body fat metabolism. Leptin is a product of ob gene created by adipocytes. It seems to signal metabolic information to the reproductive system. Objective: The aim of this study to evaluate the relationship between serum leptin and infertility in oligozoospermic men. Materials and Methods: Seventy men were investigated; fertile normozoospermic as a control (n=35) and infertile oligozoospermic (n=35). The patients underwent estimation of body weight (kg), height body mass index (BMI), semen analysis, serum FSH, LH, testosterone and leptin, lipid profile and serum malondialdehyde (MDA). The infertile group was given a treatment course of clomiphene citrate (50 mg) and vitamin E (400mg) and then all the previous parameters were reevaluated after 3 months. Results: Mean body weight was significantly higher in infertile oligozoospermia compared to controls. The height showed no significant difference between the groups. Hormonal profile revealed significant (P=0.0001) difference in FSH between control group and oligozoospermic group (7.04±4.20 versus 3.35±1.47 mIU/ml), but after treatment with clomiphene citrate and vitamin E there is a slight non-significant (P=0.231) decrease in the level of FSH in oligozoospermic group. The level of testosterone showed a statistically significant (P=0.0001) difference between fertile normozoospermic (5.52 ±1.29 ng/ml) and infertile oligozoospermic. This hormone showed a significant (P<0.0001) increase in its level after treatment with clomiphene citrate and vitamin E (3.48±1.56 ng/ml). Other hormone showed non significant difference between the two groups pre and post treatment. Lipid profile showed no significant difference between the two groups. There was no significant (P=0.477) difference in serum MDA between the control and the infertile groups (4.69±1.32 vs. 5.18±1.19 µmol/L) respectively, but after treatment the results showed a mild significant (P=0.026) decrease in comparison to the level in the control group and also a significant (P=0.0001) decrease in comparison to the level before treatment. Infertile oligozoospermic had significantly higher serum leptin level (8.03±1.22 ng/ml) than control (3.62 ± 1.00 ng/ml). Conclusion: Serum leptin demonstrated a significant positive correlation with age, body weight, BMI and a significant inverse correlation with serum testosterone. It had nonsignificant correlation with the height and sperm concentration. These results are suggestive of a link between the adipocyte derived hormone leptin and male reproduction.
The compromise of uterine receptivity is one of the main reasons for the failure of assisted reproduction techniques. Leukemia inhibitory factor is one of those molecular parameters that allow us to identify an endometrium as receptive. Platelet rich plasma (PRP) contains a range of cytokines and growth factors including leukemia inhibitory factor that encourage the migration, proliferation, and differentiation of cells. Treatments for antagonist ovarian stimulation were administered to 75 women under the age of 40, and 2-D power doppler ultrasound was used to guide the oocyte harvest. Patients were allocated into three groups according to the time of intrauterine infusion of PRP (no PRP, PRP at triggering day and PRP at ova pickup day). Power Doppler is utilized to measure endometrial thickness (ET), pulsatility index (PI), and resistance index (RI) of sub endometrial arteries, as well as serum levels of leukemia inhibitory factor (LIF). The mean endometrial thickness as well as serum level of LIF in PRP trigger group were higher, resistance index and pulsatility index were lower in a significant manner than control and PRP ova pickup group at all the days of study (triggering, ova pickup and embryo transfer days). These findings revealed that intrauterine PRP infusion on ova pick up day is associated with a significant increase in serum level of LIF that in turn enhances endometrial thickness for patients scheduled for intracytoplasmic sperm injection (ICSI).
Metformin is an anti-hyperglycemic medication used to treat type 2 diabetes mellitus. Regardless of its anti-diabetic effect, metformin has a non-genomic action and it could be a beneficial additive to treat the sperm which considered as transcriptionally dormant cell and also a molecule that could rapidly adjust the metabolism of spermatozoa to adapt it to the surrounding environment. Sixty-four normozoospermic semen sample included in the study. Each sample incubated for 30 minutes with metformin (2.5 and 0.5 µM) and SFA was conducted to each sample. There was a significant decrease in total motility (progressive and non-progressive) in samples incubated with both concentrations compared with the neat samples. While the samples incubated with 0.5 µM shows a lower motility compared with 2.5 µM. Metformin significantly reduced total motility of fresh spermatozoa at both concentrations.
Kisspepetin is considered as a key regulator of reproduction, through its direct role in controlling the hypothalamic pituitary gonadal axis necessary for controlling the ovulatory process and indirectly by impacting glucose heamostasis and metabolism. We aimed to examin the correlation between body mass index and level of Kisspeptin in serum and follicular fluid. This study is a cross sectional study done on (80) infertile Iraqi patients aged (18-40 years) scheduled for fresh intracytoplasmic sperm injection cycle. This study is conducted in the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies of Al-Naharin University, from September 2020 to May 2021. For each participant, the GnRH antagonist protocol was used to hyperstimulate the ovaries, and samples from serum and follicular fluid were obtained at day of oocytes retrieval for measuring Kisspeptin level using enzyme linked immunosorbent assay (ELIZA) technique. Serum and follicular fluid Kisspeptin were insignificantly increasing with the increase in the BMI (p value =0.406) (p value = 0.352) respectively. Serum and follicular kisspetin were positively and significantly correlated (r = 0.536, p value= 0.000). Serum and follicular fluid kisspeptin were numerically increasing with the body mass index of infertile women, although this increament is statically insignificant, serum kisspeptin can be used as a mirror image to follicular fluid kisspeptin as there was a statistical association between them in this study.
Resistin and follistatin levels have been found to be high in polycystic ovarian syndrome (PCOS) patients, that play role in pathophysiology of PCOS and pregnancy status of women having ICSI cycle. This study aimed to determine the relationship between the hormone levels and pregnancy outcome in PCOS and non-PCOS women undergoing intracytoplasmic sperm injection (ICSI) program. The study was done at Higher Institute for Infertility Diagnosis and Assisted Reproductive Technologies from 2022 to 2023. Eighty infertile women participating in the IVF/ICSI treatment. They were split to control (40 non-polycystic ovarian syndrome-PCOS women) and 40 women for PCOS group. They were subdividing to subgroups’ according to body max index (BMI; normal and overweight / obese). ELISA method was used to measure hormone level at day 2 and follicular fluid and serum at day of ova pick up. About 38.8% of study participants were got pregnant. Means of serum resistin and follistatin in day 2 and OPU were significantly higher in patients with PCOS and normal BMI level while follistatin at day 2 and in follicular fluid (FF) were significantly higher in participants who got pregnant. Follistatin level at day 2 > 9.89 ng/ml and in FF > 15.06 ng/ml and might be as a predictor for occurrence of pregnancy after ICSI. The cutoff point of follistatin level in the serum at day two and in FF at the day OPU was found to be a predictor of pregnancy following ICSI, while no correlation was found between resistin and pregnancy outcome.