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: 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: 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: 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.
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.
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).
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.
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.
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.
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: 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: 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 : 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: 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: 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: 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.
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: 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.
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 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: 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.
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: 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: 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.
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: 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.
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.
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 .
Male infertility is a global health issue affecting over 20 million males. A spermatogenesis-related cell differentiation abnormality causes teratozoospermia (TZS). Extensive research has linked TZS to poor fertilization outcomes at assisted reproductive technology (ART) centers as well as to spontaneous pregnancies. This study used Glycyrrhiza glabra (Gg) and Platelet Rich Plasma (PRP) media to establish a culture medium that aimed to improve certain sperm functions of teratozoospermic men in vitro. A total of fifty semen samples were examined. The World Health Organization 2021 and 1999 standards were used to evaluate certain sperm parameters and the DNA fragmentation index. Platelet-rich plasma and Gg extract were utilized as a medium for in vitro preparation and activation of semen samples complaining of teratozoospermia factor. Following activation by the utilization of the density gradient approach and the combination of Gg and PRP medium, there was a statistically significant ( p <0.001) improvement in certain sperm function metrics. These improvements included a higher percentage of morphologically normal sperm, a rise in active motility, and a drop in the DNA fragmentation index. The density gradient technique, regardless of the medium used in this investigation, resulted in a significant enhancement of morphologically normal sperms with other certain sperm parameters and a decrease in the DNA fragmentation index.
Background: The sperm plasma membrane activity and absence of antisperm antibodies in infertile patients ejaculate significantly associated with fertilization potential of human spermatozoa. The sperm viability detected by hypoosmotic swelling test (HOST) and autoimmune infertility detected by antisperm antibody assay presented diagnostic and prognostic tools for male infertility factors. Objective: This study was devised to compare the results of hypoosmotic swelling test and antisperm antibody assay for normozoospermic men and infertile patients affected by autoimmune infertility. Methods: Thirty semen samples were collected by masturbation and prepared by direct layering technique. Hypo-osmotic swelling test was performed by mixing 0.1 ml of semen with 0.9 ml of a 150 mOsm/L NaCl as a hypo-osmotic solution and direct immunobead assay were used to determine the presence of Antisperm Antibodies (ASA) bound on sperm surface. However, sperm concentration, sperm motility, and normal sperm morphology were evaluated according to World Health Organization (WHO) criteria and subjected to HOS test and ASA assay. The sperm prepared and incubated for 30 minute in 5% CO2 at 37°C after in vitro sperm preparation. Results: The results of the present study reported that the percentage of HOS test score in antisperm antibodies positive sperm samples (52.30 ±5.2) was significantly lower than that noticed in antisperm antibodies negative (70.30 ± 3.4) sperm samples. Conclusion: It was concluded that the use of HOS test as a simple and diagnostic and prognostic tools for identify male infertility. Further studies are suggested to assess the effect of ASA on sperm plasma functional integrity after ICSI and in vitro fertilization and embryo transfers (IVF-ET).
Background: 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.
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: 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: 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.
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: 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.
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: Tribulus terrestris (TT), Phoenix dactilyfera (Pd) and Nasturtium officinale (No) were believed to have fertility-enhancing effects and given by herbalists in Baghdad to men who need to increase their fertility capacity. Objectives: To investigate the possibility that aqueous and ethanolic extracts of the plants mixture can counteract the adverse effect of theobromine on reproductive system of male mic. Materials and Methods: One hundred and twenty mature male mice were injected intraperitoneally with 250 mg/kg/day of theobromine for four weeks. Then the males were treated intraperitonially with different dose levels of aqueous and ethanolic extracts of a mixture of the three plants for four weeks to evaluate their effects on body weight, reproductive organs weight, sperm parameters, hormones level and reproductive performance. Results: Both extracts showed a significant enhancement in all parameters studied. Almost all these results showed a dose dependent pattem with the ethanolic extract being the more effective than aqueous extract in all parameters. Conclusions: Mixture of the three plants seems to counteract the action of theobromine on the reproductive system of male mice through enhancing the concentration and motility of spermatozoa, increasing epididymis and seminal vesicles weight which are an indicator of testosterone production which brought about the stimulation of all male reproductive organs.
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: 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: 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.
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, 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: 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 .
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: 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.
Oligoasthenoteratozoospermia is a prevalent cause of male infertility. (iOAT). It is generally acknowledged that, due to the fact that spermatogenesis is a sex-hormone-dependent process, specific endocrine treatments might enhance or recover male fertility. The typical negative feedback of sex steroids is disrupted by anti-estrogen medication, leading to a rise in endogenous gonadotropin-releasing hormone secretion (FSH and LH) straight from the pituitary. The stimulation of Leydig cells in the testes by FSH and LH is thought to promote local testosterone synthesis, which in turn may improve spermatogenesis. The study sample consisted of 42 iOAT patients. Baseline seminal fluid analysis (SFA) and hormonal assessment (FSH, LH, prolactin, and testosterone) were taken. Each patient was given medical treatment with tamoxifen (20 mg/day) for three months. SFA, FSH, and testosterone were taken after three months. There was a significant increase in FSH, testosterone, sperm concentration, and progressive motility after treatment (p value <0.001), furthermore, there was no significant correlation between hormonal changes and changes in SFA study markers (p value > 0,05). Improvements in hormone levels following tamoxifen treatment for patients with idiopathic oligoasthenoteratospermia do not correlate with alterations in SFA markers.
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.