Background: Over 70 million couples suffer from infertility worldwide and the majority of those couples can be found in developing countries. Aims of study: This study aims to determine if there is any significant difference in levels of inhibin B and gonadal hormones in fertile and infertile men. Subjects, Materials & Methods: Clinical and hormonal characteristics related to male infertility factor of the infertile patients(n=75)and representative proven fathers as a control(normozoospermic ones)(n=25)were studied. All males attending infertility clinic at Baquba General Hospital, Diyala in Iraq. The study included the measurement of gonadal hormones levels by using (ELISA)method and it was also used in analyzing the inhibin B hormone. The VIDAS instruments were used for enzyme immunoassay measurements Luteininzing Hormone(LH),Testosterone(T) ;and Follicle Stimulating Hormone(FSH). The samples were analyzed and classified according to WHO criteria. Results: The results of this study showed that inhibin B hormone and other gonadal hormones in control (representative proven fathers-normozoospermic ones)as inhibin B (240.2±17.4)Pg/ml, LH (1.639±0.103)mIU/ml, Testosterone (0.6253±0.035)ng/ml and FSH (0.469±0.0103)mIU/ml. In asthenozoospermic ones as an example of infertile patients the results showed that inhibin B (76.9±8.99)Pg/ml, LH(1.4311±0.033)mIU/ml, T (0.5483±0.030)ng/ml and FSH (0.495±0.0124)mIU/ml. Except the inhibin B comparisons all of other non significant evaluated parameters & correlations within this study could be ascribed to the role of cascaded role of inhibin B with the gonadal hormones in spermatogenesis. Conclusions: It concluded from this study that decreased levels of inhibin B, LH,Testosterone, but increased levels of FSH hormones were shown in asthenozoospermia group as an example of infertile patients,and increased levels of inhibin B,LH and Testosterone but decreased levels of FSH were shown in normozoospermia as control.
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.
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: Immunological infertility is expected to be the reason of infer-tility in 9-36% of the concerned twosomes. The major cause of immunological infertility is the creation of anti-sperm antibodies (ASA), which influence the capability of insemination of spermatozoa. It has been detected that anti-sperm antibodies (ASA) are present either systemically in blood or locally in seminal plasma of approximately 10% of infertile male patients. Immunity to sperm can cause infertility; humoral antibodies directed against sperm did not necessarily impair fertility unless the circulating antibodies are also present within the re-productive tract and on the living sperm surface. Hormonal imbalance and sperm autoimmunity have been considered as two systems that play in near association and affect each other. Testosterone is the Steroid hormone necessary for the de-velopment and maintenance of secondary sexual characteristics as well as initi-ation and maintenance of Spermatogenesis .It was known that males with abnor-mal seminal fluids have lower concentrations of the testosterone hormone. Objectives: To study the serum & seminal plasma antisperm antibodies levels in relevance to levels of testosterone in serum and seminal plasma accordingly to sperm function parameters in different groups of infertile patients. Subjects,Materials and Methods: Blood and semen samples were collected from (80) subjects (60) infertile patients (20Asthenozoospermic, 20 Oligozoospermic,20Azoospermic) and 20 Normozoospermic subjects .The anti-sperm antibody and testosterone levels were measured by using Enzyme Linked Immunosorbent Assay (ELISA). Results: The Antisperm antibodies were positive in serum of normozoospermic, asthenozo-ospermic, oligozoospermic and azoospermic men respectively are about 10%, 5%, 20%,5% and in the total group was 10% positive. The ASA in seminal plasma of normozoospermic men, asthenozoospermic men, oligozoospermic men and azoospermic men respectively are 0%, 0%, 5%, 5% and in the total groups 2.5%.These findings lead to no possibility to make clear correlations with the studied hormones. There was significant increase (P<0.05) in the levels of testosterone in serum and seminal plasma in comparison between normozoospermic and infertile men subgroups. There was significant decrease (P<0.05) in the levels of pro-gesterone in serum and seminal plasma in comparison between normozoospermic men and infertile men subgroups, the highest levels were observed in asthenozoospermic subgroup. The study showed that 10 patients (12.5%) out of 80 patients showed a positive ASA positive in serum or seminal plasma of all groups. The result was considered positive if the value was ≥60 RU / ml .On these basis and regarding that screening criteria the positive ASA group showed the association with the low levels of testosterone in both types of the samples(serum and seminal plasma), these results was exhibited a high significant difference when compared with negative ASA group. Conclusion: Determination of the relevance of the levels of testosterone and anti-sperm anti-bodies in the serum and seminal plasma, which in turn is important to determine the type of infertility especially the immunological type.
Background: All men have the natural desire to have their own biological children. Cryopreservation of male gametes is an important aspect of human fertility preservation. With the advancement in assisted reproductive technology, indication for sperm cryopreservation is expanding. Objectives: To evaluate the effect of cryopreservation by using Sperm Freeze TM kit on some sperm parameters for infertile patients. Materials and methods: Ninety five semen samples were taken from male patients. The seminal fluid analysis was performed according to the standard criteria of WHO (1999). The patients were divided into four groups depending on their sperm concentration and motility: Normozoospermic, oligozoospermic, asthenozoospermic and oligoasthenozoospermic patients. Cryopreservation in liquid nitrogen was done by using Sperm Freeze TM kit for the samples and thawing was performed after three months. Results: In general, the post-thawing study revealed a considerable highly significant (P <0.001) decrease in the sperm parameters which include the means of sperm concentration, sperm motility, progressive motility and normal sperm morphology as compared to that of pre-cryopreservation. Whereas the percentage of immotile sperm was highly significant (p<0.05) increase as compared to pre-cryopreservation in all groups except for oligozoospermic patients in which there was a significant (p<0.05) increase. Conclusions: After semen cryopreservation, all parameters were significantly decreased.
Human 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 : With the rapid evolution of assisted reproductive technologies, more and more studies are concerned about the possible influential factors on the success rates of these procedures, with the efforts are now mainly directed toward defining these factors in a trial to increase these rates. Of these factors, inflammation is considered among the most remarkable and is in the focus of the updated studies. However, most of these studies are evaluating the role of inflammation in patients with polycystic ovarian syndrome(PCOS) , while the impact of this inflammation in insulin resistant non PCOS patients is underesti mated and not well evaluated . Objectives: This study is a trial to elucidate the importance of one of the inflam matory mediators , Tumor Necrosis Factor-alpha (TNF – α) , in insulin resistant patients undergoing IVF ( ICSI ) cycles . Patients and Methods: A group of 27non-PCOS infertile patients were select ed for this clinical randomized prospective study from patients attending the high institute for infertility diagnosis and assisted reproductive technologies / Al-Nahrain University and subjected to intra-cytoplasmic sperm injection (ICSI) procedure. BMI was measured for all patients and they were classified according to WHO classification into 3 groups : Acceptable , overweight and obese . Fast ing plasma level of glucose and insulin was investigated and insulin resistance index (HOMA) was calculated. Serum TNF-α has been measured using ELISA and the relation with insulin resistance was investigated. Finally, the pregnancy rate, as a primary reproductive endpoint was compared according to TNF -a and insulin resistance levels between different study subjects. Results: Of the 27 infertile patients , 17(63 %) were non obese and 10 (37%) were obese , 15(55.5%) were having HOMA index < 1.6 and 12 (44.5 %) were having HOMA index ≥ 1.6 . BMI was found to be positively correlated with high er HOMA index ( P = 0.01 and R= 0.4 in spearman correlation). Investigating the relation between TNF-α with BMI revealed that TNF-α levels are increasing with the increment in BMI, However , the difference in the mean TNF-α between the three BMI groups failed to achieve statistical significance (P=0.1). In contrast to that, when stratifying for HOMA index , TNF-α was significantly correlated with it ( spearman correlation R= 0.4 , P=0.01 ) , and patients with HOMA index ≥1.6 were found to have higher levels of TNF-α ( P = 0.03) and to be more obese ( P=0.02 ) than those with HOMA index <1.6.
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: The intra uterine insemination (IUI) still an easy, simple and effective method to overcome the infertility that resulted from different causes. Although the equipments and methods of assisted reproductive technologies (ART) were highly developed, the percentage of successful IUI live-birth not exceed 20-30%. Objective: The present study was aimed to elucidated the cut off values of in vitro sperm preparation technique, follicular number and size, endometrial thickness and hormonal profile that lead to successful pregnancy rate (PR)following IUI. Materials and Methods: In retrospective study, one hundred cases of infertile couples were in who were became pregnant following IUI at the Institute of Infertility Diagnosis and Assisted Reproductive Technologies, through the period from January 2007 to January 2010. Depending on the results of retrospective study, IUI was achieved for 100 infertile patients between August 2010and June 2011. The mean of significant prognostic variables were measured in both studies namely: age, type of infertility, type of sperm activation techniques, the protocol of ovulation induction medicine, the mean of hormonal status of Luteinizing Hormone (LH), Estradiol(E2), and number with diameter of follicles, endometrial thickness a cycle day before hCG injection. Results: The protocol of ovulation induction in IUI cycles by using clomiphene citrate(CC) and recombinant gonadotropin (rFSH) was significantly(p<0.05) improve the IUI results compared to CC alone or CC with PergonalS. The mean values obtained by ultrasonography namely; two follicles, and dominant follicle of 19.5mm in size with endometrium thickness of >9.5mm were gave significantly successful IUI outcome compared to other values. In prospective study similar results were obtained, therefore the overall pregnancy rate was 37% per cycle. Conclusion: The calculated data of successful IUI outcome from retrospective study gave a best pregnancy rate in prospective study which was 37%. Consequently, these results can be considered as guideline for physician who interest in assisted reproduction to have a success in IUI.
Background: 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: 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: 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 : 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.