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: 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: 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: 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.
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.
Human peptidoglycan recognition protein-1 (PGLYRP‐1) protein molecule activation in the semen of infertile patients has been suggested to induce a negative impact on seminal fluid analysis. This study aimed to evaluate Human Peptidoglycan Recognition Protein 1 for indication of bad seminal fluid analysis parameters. One hundred seventy-six infertile patients were included in this study. Seminal fluid analysis and Endtz test were done; Human Peptidoglycan Recognition Protein 1 in seminal plasma using enzyme-linked immune-sorbent assay technology PGLYRP‐1 is evident in seminal plasma of infertile men and statistically different according to the type of infertility. The concentration of PGLYRP-1 in the semen is directly proportional to the presence of leukocytes (Endtz test), which is reflected negatively on the quality of the semen. To the best of our knowledge, this study has not been done in Iraq or elsewhere. The presence of PGLYRP-1 protein in the semen may be evidence of leukocytospermia that may be related to bad quality for semen across infertile men with higher sensitivity than the Endtz test.
Background : 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.
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: 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.
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.
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.
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: 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: 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.
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.
Phosphatidylserine (PS), exposed on viable sperm, is required for fertilization as it is recognized by specific receptors located on the microvilli of the oocyte to promote sperm: egg fusion. The heads of viable and motile sperm exhibit PS. With (PS) exposure progressively increasing during sperm transit through the epididymis. PS asymmetry on the leaflets of plasma membrane is arranged in a complex way, via multiple proteins functioning in opposite directions; further, PS is also essential for many intracellular trafficking events. The aim of this study was to evaluate the phosphatidylserine PS level in seminal plasma of infertile men before and after activation using Direct Swim Up (DSU) and Indirect Swim Up (IDSU) activation method and make a comparison between them. Forty infertile men were included in this study during their attendance to the infertility clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University. Normozoospermic or mild asthenozoospermic semen samples were used to choose the cases, according to WHO manual standards (1999 -2010). The collected semen samples obtained were divided into three parts: The first part prepared was as an in vitro sperm characterization before activation while the second and third parts were after activation using direct and indirect swim up techniques respectively. The level of seminal plasma phosphatidylserine was increased after the activation techniques from the baseline level before activation. Comparison of Seminal plasma phosphatidylserine levels between direct and indirect sperm activation technique were significantly higher in direct swim up activation technique than indirect one. Phosphatidylserine (PS) in semen plasma of infertile men was significantly increased after activation using Direct Swim Up DSU preparation method.
Background: 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.
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.
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: 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: 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.
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: The immune infertility caused by anti-sperm antibodies (ASAs) represented about 1020%- of infertility among the couples, the ASAs interfere with sperm parameters such as the sperm motility and sperm ability to penetrate cervical mucus, sperm-oocyte binding, and fertilization and embryo developments. Objectives: The present study designed to assess semen analysis, presence of ASAs and DNA fragmentation index as well as correlation within these parameters in normzoospermic Iraqi subjects Patients, Materials and Methods: A total number of Iraqi subjects (116) with range of age (2051-) years and their mean duration of infertility (4.70 ± 2.77). Seminal fluid for Macroscopic and Microscopic assessments done according to WHO 2010 criteria. The mixed agglutination reaction (MAR) test used to assess theirASAs in semen (direct method), seminal plasma and serum (indirect method); for the both IgG and IgA classes of antibodies and their distribution among different parts of spermatozoa as percentage. Results: The mean of IgG using direct method was (23.881.75±) and for indirect detection method the mean was (27.412.41±) with no significant difference (P<0.05) between the two methods. While the direct method for IgA detection mean (14.46 ±1.76) and the mean for indirect method of detection (6.860.39±) and there was significant difference (P>0.05) between the two methods. In addition to that, the distribution of IgG and IgA detected by direct method on sperm parts showed no significance difference except for the sperm tail; while the indirect method for IgA and IgG showed significant difference in distribution on sperm mid-piece. Conclusion: The correlation among sperm parameters and Immunoglobulin using direct and indirect methods showing variations in significance also in the correlation between the direct and indirect methods according to distributions on sperm surface
The 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.
Background: The Fibroblast associated (Fas) system in the testes has been identified as a key regulator of apoptosis, a process that greatly influences the germ cell population of the testes. Objective: This study measures the level of soluble Fas Ligand (sFasL) in the semen of oligozoospermic men evaluating the association between seminal plasma sFasL and spermatogenesis. Methods: A total 58 oligozoospermic men and 29 normal volunteers were included in this study. semen was evaluated according to World Health Organization 2010 standard parameters. sFasL was measured using ELISA enzyme immunoassay for quantitative determination of sFasL. Results: s FasL level was found significantly higher (P< 0.01) in seminal plasma of oligozoospermic men, with weak correlation of the level of this marker with the degree of severity of oligozoospermia. Conclusions: The apoptotic marker, sFasL is a novel marker found in the seminal plasma of oligozoospermic men. It's level is higher in cases of oligozoospermia regardless of its severity.
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).
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.
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: 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.
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 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.
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.
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: 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: 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: 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.
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.
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.
Many men have reported erectile dysfunction as a consequence of COVID-19 infection. The underlying causes, whether related to hormonal changes or endothelial dysfunction, are not yet fully understood. This study aims to investigate the impact of COVID-19 on male sexual function and fertility, specifically focusing on erectile dysfunction. A cohort of 50 males who had no prior history of erectile dysfunction before contracting COVID-19 was assessed. The study analyzes the progression of these conditions over a 90-day period post-recovery from COVID-19, with a particular emphasis on age-related factors. All males were confirmed infected with COVID-19. Comprehensive medical histories were obtained from all participants, and physical examinations were conducted. The International Index of Erectile Function (IIEF-5) questionnaire was employed to assess erectile dysfunction. Assessments were performed at three separate intervals, 30, 60, and 90 days after their recovery from COVID-19. Participants consistently reported varying degrees of erectile dysfunction, which appeared to improve over time. Notably, the participants who reported erectile dysfunction tended to be older than those who did not, and this age-based distinction was highly significant (p = 0.004). In conclusion, this study highlights the impact of COVID-19 infection on male sexual function, with age and the time post-recovery emerging as significant factors contributing to these effects.
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.
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.
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.
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.
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.
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).
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.
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).
Intrauterine insemination (IUI) comprises the insertion of a high number of washed and processed spermatozoa directly into the cervix to the uterine cavity around the time of ovulation to increase the chances of pregnancy. It is usually used for couples with mild male factor infertility, unexplained infertility, cervical factors, mild endometriosis, and psychological, sexual dysfunction. The endometrium, which lines the uterus, is prepared for the implantation of the embryo; this process starts in the proliferative phase and extends throughout the luteal phase. Implantation, a crucial phase of pregnancy, is a complicated process, and more than half of failures to achieve pregnancy are caused by implantation failure. To assess the influence of endometrial flushing of 1000IU hCG done prior to IUI procedure on pregnancy rate in infertile females undergoing IUI treatment. The present study was applied to 80 infertile subjected to ovarian stimulation protocol using letrozole and hMG in doses adjusted according to their response; triggering was done using subcutaneous HCG. The results of the present study showed no significant difference regarding initial characteristics, indication for IUI, basal menstrual cycle characteristics, ovarian stimulation characteristics, and semen parameters before and after processing, while there was a significant difference between both groups regarding biochemical and clinical pregnancy rates. In IUI cycles, endometrial human chorionic gonadotropin (hCG) flushing done 5-10 minutes prior to IUI can improve biochemical and clinical pregnancy rates.
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.