Comparison of Embryological Results of Microinjection in Two Groups of Men with and without Requesting Sperm DNA Fragmentation Index Measurement.

Introduction: The sperm DNA fragmentation index (DFI) is considered a valuable measure to assess male fertility, but the predictive value of DFI for the outcomes in assisted reproductive technology (ART) is still controversial. Therefore, this study is aimed at investigating the effect of requesting a DFI test or performing ART without DFI on the results observed in the embryology laboratory (number of embryos, fertilization rate, and embryo quality) after intracytoplasmic sperm injection (ICSI). Methods: This retrospective study was conducted on infertile men who underwent ICSI and were referred to the Avicenna Infertility and Recurrent Abortion Treatment Center in Tehran from 2019 to 2022. The samples were categorized into two groups: a case group with DFI measurement and a control group without DFI measurement. We conducted a comparative analysis of the embryology results between the two groups, focusing on parameters such as fertilization rate, number of embryos, and embryo quality. t-tests and Mann-Whitney U tests were used to conduct single variable analysis. Potential confounding effects were adjusted to use the multivariate linear and logistic regression. Results: Data analysis showed no significant statistical difference between the case group and the control group in terms of the number of embryos (95% confidence interval for the regression coefficient (ß) = -0.257-0.123), and embryo quality (95% confidence interval for ß = -0.199-0.114). There was no significant statistical difference between the two groups due to the fertilization rate (95% confidence interval for ß = -3.42-3.42), except for the variables of woman's age and sperm count after ICSI, as determined by adjusted linear regression. Conclusions: Although DFI measurement is used to assess male infertility, its importance as a predictor for the embryology outcomes after ICSI requires further evaluation and the determination of a cut-off point for predicting results. This study was based on retrospectively collected DFI data, and prospective studies confirming the superiority of ICSI outcomes are necessary.

Targeted Amino Acids Profiling of Human Seminal Plasma from Teratozoospermia Patients Using LC-MS/MS.

Identifying the metabolome of human seminal plasma (HSP) is a new research area to screen putative biomarkers of infertility. This case-control study was performed on HSP specimens of 15 infertile patients with teratozoospermia (defined as normal sperm morphology < 4%) and 12 confirmed fertile normozoospermic men as the control group to investigate the seminal metabolic signature and whether there are differences in the metabolome between two groups. HSPs were subjected to LC-MS-MS analysis. MetaboAnalyst5.0 software was utilized for statistical analysis. Different univariate and multivariate analyses were used, including T-tests, fold change analysis, random forest (RF), and metabolite set enrichment analysis (MSEA). Teratozoospermic samples contained seventeen significantly different amino acids. Upregulated metabolites include glutamine, asparagine, and glycylproline, whereas downregulated metabolites include cysteine, γ-aminobutyric acid, histidine, hydroxylysine, hydroxyproline, glycine, proline, methionine, ornithine, tryptophan, aspartic acid, argininosuccinic acid, α-aminoadipic acid, and ß-aminoisobutyric acid. RF algorithm defined a set of 15 metabolites that constitute the significant features of teratozoospermia. In particular, increased glutamine, asparagine, and decreased cysteine, tryptophan, glycine, and valine were strong predictors of teratozoospemia. The most affected metabolic pathways in teratozoospermic men are the aminoacyl-tRNA, arginine, valine-leucine, and isoleucine biosynthesis. Altered metabolites detected in teratozoospermia were responsible for various roles in sperm functions that classified into four subgroups as follows: related metabolites to antioxidant function, energy production, sperm function, and spermatogenesis. The altered amino acid metabolome identified in this study may be related to the etiology of teratozoospermia, and may provide novel insight into potential biomarkers of male infertility for therapeutic targets.

The Amino Acid Profile in Seminal Plasma of Normozoospermic Men: A Correlation Analysis with Spermiogram Parameters and Total Antioxidant Capacity.

Background: Male infertility is usually determined by the manual evaluation of the semen, namely the standard semen analysis. It is currently impossible to predict sperm fertilizing ability based on the semen analysis alone. Therefore, a more sensitive and selective diagnosis tool is required. Methods: Twelve fresh semen samples were collected from fertile volunteers attending the Avicenna Fertility Center (Tehran, Iran). The seminal plasma (SP) was prepared and subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the total antioxidant capacity (TAC) was analysis. Thirty-four amino acids including essential amino acids (EAA), non-essential amino acids (NEAA), and non-proteinogenic amino acids (NPAA) relative concentration were determined, and the correlation between their concentration with spermiogram parameters and TAC of the SP was analyzed. Results: Significant positive correlations have been found between selected amino acids with the motility (Met and Gln, rs=0.92; Cys, rs=0.72; and Asn, rs=0.82), normal sperm morphology (Met, rs=0.92; Cys, rs=0.72; Glu, rs=0.92; and Asn, rs=0.82), and sperm concentration (Trp, Phe, and Ala). In contrast, several AAs, including Gly, Ser, and Ile showed negative correlations with sperm concentration (rs=-0.93, r=-0.92, and r=-0.89, respectively). Furthermore, TAC showed a positive association only with Tyr (rs=0.79). Conclusion: The strong positive/negative correlations between the seminal metabolic signature and spermiogram demonstrate the significance of determining metabolite levels under normal conditions for normal sperm functions. Combining the metabolome with the clinical characteristics of semen would enable clinicians to look beyond biomarkers toward the clinical interpretation of seminal parameters to explain the biological basis of sperm pathology.

Sperm Retrieval in Non-azoospermic Patients with Persistent Ejaculation Dysfunction.

Infertility is a common disease that affects 15 to 20% of couples at some point in their lives. Among infertile couples, male factor accounts for 50% of infertile cases. Assisted reproductive techniques are the gold standard approach in case of failure in medical or surgical treatments. Moreover, the role of the urologist in these approaches is to provide appropriate sperm on the day of oocyte pick-up. However, sperm retrieval procedure is quite different in azoospermic and non-azoospermic men. Although most cases of infertile patients are not azoospermic, their ejaculation disorder prevents obtaining sperm for assisted reproductive techniques. This review article explains common problems of sperm retrieval in non-azoospermic patients with persistent ejaculatory dysfunction and introduces some management strategies. In fact, it is possible to design a classic approach for managing such patients, which definitely reduces the problems faced by clinicians as well.

The mouse testis tissue culture could resume spermatogenesis as same as in vivo condition after human spermatogonial stem cells transplantation.

OBJECTIVE: The introduction of alternative systems in vivo is very important for cancer patients who are treated with gonadotoxic treatment. In this study, we examine the progression of the spermatogenesis process after human spermatogonial stem cell (SSCs) transplantation in vivo and in tissue culture conditions. MATERIALS AND METHODS: Human SSCs were obtained from a Testicular Sperm Extractions (TESE) sample, and characterization of these cells was confirmed by detecting the promyelocytic leukemia zinc finger (PLZF) protein. These cells, after being labeled with Di-alkyl Indocarbocyanine (DiI), were transplanted to adult azoospermia mouse testes treated with Busulfan 40mg/kg. The host testicular tissue culture was then considered a test group and in vivo transplant a control group. After 8 weeks, immunohistochemical, morphometric and molecular studies were performed. RESULTS: The results of morphometric studies indicated that the mean number of spermatogonia, spermatocytes, and spermatids in the test groups was significantly lower than in the control group (P<0.05) and most of the cells responded positively to DiI tracing. Immunohistochemical study in both groups revealed expression of PLZF, Synaptonemal complex protein 3 (SCP3) and Acrosin Binding Protein (ACRBP) proteins in spermatogonial cells, spermatocyte and spermatozoa, respectively. Also, PLZF, Transition Protein 1 (TP1) and Tektin-1 (Tekt1) human-specific genes had a significant difference in the between test groups and control groups (P<0.05) in molecular studies. CONCLUSION: These results suggest that the conditions of testicular tissue culture after transplantation of SSCs can support spermatogenesis resumption, as well as in an in vivo condition.

Effects of coadministration of DHA and vitamin E on spermatogram, seminal oxidative stress, and sperm phospholipids in asthenozoospermic men: a randomized controlled trial.

BACKGROUND: It is unknown which compounds in spermatozoa or seminal plasma may be involved in the regulation of sperm motility. OBJECTIVES: The aim of this study was to investigate the effects of DHA (22:6n-3), vitamin E, and their probable interactions in men with asthenozoospermia. METHODS: A factorial, randomized, double-blind, placebo-controlled trial was conducted in infertility clinics in Tehran, Iran. The participants were idiopathic asthenozoospermic men aged 20-45 y, with normal endocrine function. Their concentration of spermatozoa and percentage of morphologically normal spermatozoa were equal to or above the lower reference limits, according to the fifth edition of the WHO guideline. Out of 717 men referred to the infertility clinics, 180 asthenozoospermic men were randomly assigned to 1 of 4 groups according to stratified blocked randomization by age and sperm concentration. Participants took daily 465 mg DHA plus 600 IU vitamin E (DE), 465 mg DHA plus placebo (DP), 600 IU vitamin E plus placebo (EP), or both placebo capsules (PP) for 12 wk. Sperm characteristics, oxidative stress of seminal plasma, serum and sperm membrane fatty acids, dietary intakes, anthropometric measurements, and physical activity were measured at baseline and after 12 wk. RESULTS: After the intervention, mean ± SD sperm progressive motility was greater in the DE group (27.9 ± 2.8) than in the DP (25.7 ± 3.4), EP (26.1 ± 2.8), and PP (25.8 ± 2.6) groups (P < 0.05). Sperm count (P = 0.001) and concentration (P = 0.044) increased significantly in the DE group compared with the other 3 groups, whereas other semen parameters were not significantly different between the groups after the intervention. Serum concentrations of n-3 PUFAs were significantly higher in the DE and DP groups than in the EP and PP groups. CONCLUSIONS: Combined DHA and vitamin E supplements led to increased sperm motility; however, no significant changes occurred in sperm morphology and vitality in asthenozoospermic men.This trial was registered at clinicaltrials.gov as NCT01846325.

In vitro transplantation of spermatogonial stem cells isolated from human frozen-thawed testis tissue can induce spermatogenesis under 3-dimensional tissue culture conditions.

BACKGROUND: Sperm production is one of the most complex biological processes in the body. In vitro production of sperm is one of the most important goals of researches in the field of male infertility treatment, which is very important in male cancer patients treated with gonadotoxic methods and drugs. In this study, we examine the progression of spermatogenesis after transplantation of spermatogonial stem cells under conditions of testicular tissue culture. RESULTS: Testicular tissue samples from azoospermic patients were obtained and then these were freeze-thawed. Spermatogonial stem cells were isolated by two enzymatic digestion steps and the identification of these cells was confirmed by detecting the PLZF protein. These cells, after being labeled with DiI, were transplanted in azoospermia adult mice model. The host testes were placed on agarose gel as tissue culture system. After 8 weeks, histomorphometric, immunohistochemical and molecular studies were performed. The results of histomorphometric studies showed that the mean number of spermatogonial cells, spermatocytes and spermatids in the experimental group was significantly more than the control group (without transplantation) (P < 0.05) and most of the cells responded positively to the detection of DiI. Immunohistochemical studies in host testes fragments in the experimental group express the PLZF, SCP3 and ACRBP proteins in spermatogonial cells, spermatocyte and spermatozoa, respectively, which confirmed the human nature of these cells. Also, in molecular studies of PLZF, Tekt1 and TP1, the results indicated that the genes were positive in the test group, while not in the control group. CONCLUSION: These results suggest that the slow freezing of SSCs can support the induction of spermatogenesis to produce haploid cells under the 3-dimensional testicular tissue culture.

Successful Human Spermatogonial Stem Cells Homing in Recipient Mouse Testis after In Vitro Transplantation and Organ Culture.

OBJECTIVE: In vitro transplantation (IVT) of spermatogonial stem cells (SSCs) is one of the most recent methods in transplantation in recent decades. In this study, IVT and SSCs homing on seminiferous tubules of host testis in organ culture have been studied. MATERIALS AND METHODS: In this experimental study, human SSCs were isolated and their identities were confirmed by tracking their promyelocytic leukemia zinc finger (PLZF) protein. These cells were transplanted to adult azoospermia mouse testes using two methods, namely, IVT and in vivo transplantation as transplantation groups, and testes without transplantation of cells were assigned in the control group. Then histomorphometric, immunohistochemical and molecular studies were done after 2 weeks. RESULTS: After two weeks, histomorphometric studies revealed that the number of subsided spermatogonial cells (SCs) and the percentage of tubules with subsided SCs in IVT and in vivo groups were significantly more than those in the control group (P<0.05). Immunohistochemical studies in the transplantation groups confirmed that the PLZF protein was expressed in the cells subsided on the seminiferous tubule. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) demonstrated that the PLZF gene expression was only positive in the transplantation groups, but it was not significantly different between the IVT group and the in vivo group (P>0.05). CONCLUSION: Testicular tissue culture conditions after SSC transplantation can help these cells subside on the seminiferous tubule basement membrane.

In vitro transplantation of spermatogonial stem cells isolated from human frozen-thawed testis tissue can induce spermatogenesis under 3-dimensional tissue culture conditions

BACKGROUND: Sperm production is one of the most complex biological processes in the body. In vitro production of sperm is one of the most important goals of researches in the field of male infertility treatment, which is very important in male cancer patients treated with gonadotoxic methods and drugs. In this study, we examine the progression of spermatogenesis after transplantation of spermatogonial stem cells under conditions of testicular tissue culture. RESULTS: Testicular tissue samples from azoospermic patients were obtained and then these were freeze-thawed. Spermatogonial stem cells were isolated by two enzymatic digestion steps and the identification of these cells was confirmed by detecting the PLZF protein. These cells, after being labeled with DiI, were transplanted in azoospermia adult mice model. The host testes were placed on agarose gel as tissue culture system. After 8 weeks, histomorphometric, immunohistochemical and molecular studies were performed. The results of histomorphometric studies showed that the mean number of spermatogonial cells, spermatocytes and spermatids in the experimental group was significantly more than the control group (without transplantation) (P < 0.05) and most of the cells responded positively to the detection of DiI. Immunohistochemical studies in host testes fragments in the experimental group express the PLZF, SCP3 and ACRBP proteins in spermatogonial cells, spermatocyte and spermatozoa, respectively, which confirmed the human nature of these cells. Also, in molecular studies of PLZF, Tekt1 and TP1, the results indicated that the genes were positive in the test group, while not in the control group. CONCLUSION: These results suggest that the slow freezing of SSCs can support the induction of spermatogenesis to produce haploid cells under the 3-dimensional testicular tissue culture.

Adherence to the Western Pattern Is Potentially an Unfavorable Indicator of Asthenozoospermia Risk: A Case-Control Study.

OBJECTIVE: The aim of this case-control study was to examine the relationship between dietary patterns and asthenozoospermia risk. METHODS: In total, 107 incident asthenozoospermic men and 235 age-matched controls were interviewed through the infertility clinics in Tehran, Iran, from January 2012 to November 2013. Usual dietary intakes were collected using a semiquantitative food frequency questionnaire and semen quality data were analyzed according to the fifth edition of the World Health Organization (WHO) guidelines. Dietary patterns were derived using factor analysis. The first tertile served as the reference category for regression analyses. RESULTS: In principal component analysis, 2 dietary patterns emerged: a "prudent pattern" (leafy green vegetables, yellow vegetables, other vegetables, tomatoes, fish and other seafood, fruits and natural fruit juices, legumes, whole grains, poultry, tea and coffee, low-fat dairy products, and vegetable oils) and a "Western pattern" (organ meats, red and processed meats, sugar, soft drinks and confectionary, pasta, rice and refined grains, potatoes, french fries and fast foods, high-fat dairy products, hydrogenated fats, mayonnaise and fatty sauces, and snacks). After adjustment for potential confounders, participants in the highest tertile of the prudent pattern scores had 54% lower risk of asthenozoospermia compared to those in the lowest (p for trend: 0.003). Being in the highest tertile of the Western pattern was positively associated with asthenozoospermia risk (odds ratio [OR] = 2.86; 95% confidence interval [CI], 1.83-2.97). CONCLUSIONS: Our findings suggest that adherence to the Western pattern is potentially an unfavorable indicator of asthenozoospermia risk and a diet composed mainly of plant-based foods may be associated with a reduced risk.

Dietary fatty acid intakes and asthenozoospermia: a case-control study.

OBJECTIVE: To investigate the association between dietary fatty acids intakes and asthenozoospermia. DESIGN: Case-control study. SETTING: Infertility clinics. PATIENT(S): A total of 107 men with incident asthenozoospermia and 235 age-matched controls. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Assessments of dietary intakes, semen quality, anthropometry, endocrine parameters, and demographic information. RESULT(S): According to the fully adjusted model, being in the highest tertile of total saturated fatty acids (odds ratio [OR] = 1.85, 95% confidence interval [CI] 1.24-2.96), total trans-fatty acids (OR = 2.53, 95% CI 1.54-3.92), palmitic acid (OR = 1.90, 95% CI 1.26-2.74), and stearic acid (OR = 2.13, 95% CI 1.29-3.88) was positively associated with asthenozoospermia. Whereas higher intakes of omega-3 polyunsaturated (OR = 0.68, 95% CI 0.58-0.94) and of docosahexaenoic (OR = 0.53, 95% CI 0.29-0.89) fatty acids were significantly associated with reduced odds of asthenozoospermia. CONCLUSION(S): Our findings suggest that the high intake of saturated and trans-fats was positively related to the odds of having asthenozoospermia. Conversely, inverse and dose-dependent associations were found between asthenozoospermia and intake of omega-3 polyunsaturated fatty acids. The observed associations of different types of fatty acids underline the importance of the type of fat in the etiology of asthenozoospermia.

The association of seminal plasma antioxidant levels and sperm chromatin status with genetic variants of GSTM1 and GSTP1 (Ile105Val and Ala114Val) in infertile men with oligoasthenoteratozoospermia.

In this study we aimed to examine the effects of genetic variants of GSTM1 and GSTP1 (Ile105Val and Ala114Val) on GST activity, seminal oxidative stress and sperm chromatin status in infertile men with oligoasthenoteratozoospermia (OAT). The study population (n=121) consisted of 95 infertile men with OAT and 26 controls with normozoospermia. Multiplex polymerase chain reaction (PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods were utilized to detect the aforesaid genetic variants. We measured GST activity and total antioxidant capacity (TAC) of seminal plasma by spectrophotometry. Sperm chromatin integrity and maturity were assessed using toluidine blue and chromomycin A3 (CMA3-positive sperm) staining, respectively. The analysis showed that subgroups of GSTM1 null and GSTP1 C/T+T/T genotypes in comparison with GSTM1 present and GSTP1 wild type (C/C) genotypes did not have statistically significant differences in both OAT or normozoospermic men considering sperm concentration and motility, percentage of CMA3-positive sperm, seminal plasma TAC, sperm chromatin integrity and GST activity. Thus, the findings of our study suggest that there are no significant associations between GSTM1 and GSTP1 polymorphisms and sperm parameters at conventional or at molecular levels including OS status, sperm chromatin integrity or maturity in Iranian infertile men with OAT and normozoospermia. However, these polymorphisms could be related to the fertility status of the studied population but not evaluated in this study.

Prevalence of intratubular germ cell neoplasia in cryptorchid testes of infertile men.

BACKGROUND: Cryptorchidism is a common malformation in neonates; surgery or medical treatments are applied during childhood. Untreated cryptorchid testes are in the risk of intratubular germ cell neoplasia (IGCN) and consequently invasive testicular tumors which could be shown by immunohistochemistry staining for placental like acid phosphatase (PLAP) marker. OBJECTIVE: We designed this study to know the prevalence of IGCN in untreated cryptorchid testes of infertile men, in our infertility center as a refferal center. MATERIALS AND METHODS: In this cross-sectional study we assessed H&E slides of testicular samples of 13 adult infertile patients with impalpable intra-abdominal testes seeking infertility treatment; then we stained them by PLAP marker. RESULTS: Three (23.08%) samples were positive for PLAP marker means presence of IGCN in testis. One of them showed seminoma besides IGCN. CONCLUSION: According to the results of this study and the fact that there are adult untreated cryptorchid patients in our country yet, it is suggested to pay more attention in clinical examination, assessment and follow up of such patients for malignancy screening.

Intake of food groups and idiopathic asthenozoospermia: a case-control study.

STUDY QUESTION: Is there any association between the intake of different food groups and the risk of idiopathic asthenozoospermia? SUMMARY ANSWER: A high intake of processed meat and sweets was positively associated with a higher risk of asthenozoospermia, whereas a high intake of fruits, vegetables, poultry, skim milk and sea foods was associated with a lower risk. WHAT IS KNOWN ALREADY: A high intake of lipophilic foods like meat products or milk may be negatively associated with semen quality in humans, whereas some fruits or vegetables may maintain or improve semen quality. STUDY DESIGN, SIZE, DURATION: A case-control study including 72 asthenozoospermic men and 169 normozoospermic men all from infertile couples who underwent face-to-face private interviews, from January 2011 to December 2011. PARTICIPANTS/MATERIALS, SETTING, METHODS: Semen was assessed by volume, sperm concentration, motility and morphology. Usual dietary intakes were assessed using a semi-quantitative food frequency questionnaire. Odds ratios (ORs), 95% confidence intervals (CIs) and evaluation of trends were calculated using logistic regression. The first tertile served as the reference category for regression analyses. MAIN RESULTS AND THE ROLE OF CHANCE: After adjusting for potential confounders, the risk of asthenozoospermia was significantly higher in the highest tertiles of processed meat (OR: 2.03, 95% CI: 1.70-2.44) and sweets intake (OR: 2.05, 95% CI: 1.09-2.26). Conversely, being in the highest tertile of total fruits and vegetables, the intake of dark green vegetables, skim milk, poultry and sea food intake was associated with a lower risk of asthenozoospermia (P for trend = 0.04, 0.01, 0.02, 0.03 and 0.04, respectively). LIMITATIONS, REASONS FOR CAUTION: Recall bias, selection bias and measurement bias are inevitable in this kind of study and residual confounding due to omission or imprecise measurement of important covariates remains possible. WIDER IMPLICATIONS OF THE FINDINGS: Patients with asthenozoospermia should be advised to adhere to a diet rich in fruits, vegetables, poultry, skim milk and sea foods while low in processed meat and sweets. STUDY FUNDING/COMPETING INTEREST(S): This study was financially supported by the National Nutrition and Food Technology Research Institute, Faculty of Nutrition and Food Technology, Shahid Beheshti University of Medical Science, Tehran, Iran. No conflict of interests to declare.

Discrepancy in the frequency of Y chromosome microdeletions among Iranian infertile men with azoospermia and severe oligozoospermia.

AIM: There are discrepancies in the reported frequency of Y chromosome microdeletions among Iranian infertile men. The objective of this study was to determine the frequency of Y chromosome microdeletions in an Iranian population with azoospermia and severe oligozoospermia. METHODS: Totally, 94 azoospermic and 21 severe oligozoospermic patients were screened for the presence of Y chromosome microdeletions. One hundred and five fertile men were included as a control group, as well. The screening of Yq microdeletions was performed by two multiplex PCRs using six STS markers. RESULTS: No microdeletions were detected in the men with severe oligozoospermia. In the azoospermic group 2/94 (2.13%) patients showed Y chromosome microdeletions. Among them, one patient had complete deletion of the AZFc region and the other showed complete deletion of both the AZFb and AZFc regions (AZFbc). No microdeletion was identified in the AZFa region. CONCLUSION: The estimated frequency of Y chromosome microdeletions in the present survey was lower than that of previous reports in Iranian populations.

Relationship of seminal plasma antioxidants and serum male hormones with sperm chromatin status in male factor infertility.

We explored the relationship between sperm chromatin integrity, hormone levels, seminal plasma total antioxidant capacity (TAC), and routine sperm parameters in men with male factor (MF, n = 81) and non-male factor (NMF, n = 52) infertility. Semen and blood were collected and examined from men undergoing evaluation for infertility in the Avicenna Infertility Clinic. We have examined each patient for serum hormones (LH, FSH, E2, DHEA), sperm chromatin damage, level of protamination and seminal plasma TAC. Levels of FSH, LH, sperm chromatin damage, and abnormal protamination were significantly higher in MF vs. NMF groups (p < 0.001). Sperm chromatin damage was correlated with percentage of CMA(3)- positive sperm (r = 0.64, p < 0.001) and with sperm concentration (r = -0.36, p < 0.001), motility (r = -0.21, p < 0.05), and morphologically normal spermatozoa (r = -0.29, p < 0.001). Linear regression showed sperm chromatin damage was related to percentage of CMA(3)- positive sperm (p < 0.001) in ungrouped patients. It was related to both percentage of CMA(3)- positive sperm and serum DHEA in the MF group (p < 0.001 and p < 0.05, respectively). Sperm chromatin maturity assessed by CMA(3) test was inversely related to sperm chromatin damage assessed by the toludine blue assay. Male factor infertility associated with sperm chromatin damage may be related to sperm protamination and to serum DHEA.

Effects of sperm chromatin integrity on fertilization rate and embryo quality following intracytoplasmic sperm injection.

Sperm chromatin integrity has been being recognized as an important factor in male fertility. During normal fertilization, high quality sperm with intact chromatin are selected through natural selection in journey from vagina to fallopian tube. However, using Assisted Reproductive Techniques, particularly ICSI, the natural selection is bypassed. Therefore sperm with DNA breakage have the opportunity to fertilize the egg which may lead to decreased embryo quality and implantation rate. The aim of this study was to evaluate the effects of sperm chromatin integrity on ICSI outcomes. A total of 200 semen samples were collected from couples undergoing ICSI and were analyzed according to WHO criteria. Each sample was evaluated for sperm chromatin integrity using four cytochemical assays and semen processing by swim up method. The ICSI was carried out according to a long-term pituitary down-regulation protocol. The correlation between sperm parameters, sperm chromatin integrity and ICSI outcomes (fertilization rate and embryo quality) was examined. The mean number of oocyte, fertilization rate and cleavage embryos per cycles was 7.5±5.0, 74.06%±25 and 5.4±3.6, respectively. There was not significant correlation between the results of chromatin assays (AO, AB, TB, and CMA3) and fertilization outcomes following ICSI. The fertilization rate was significantly higher for a group with less than 10% chromatin abnormality (p<0.05). Sperm chromatin integrity is essential for successful fertilization, embryo development and normal pregnancy. A protamine deficiency appeared to affect fertilization rate and embryo quality. However, the presence of confounding factors such as selection of spermatozoa according to normal morphology may influence the effect of sperm chromatin status on ICSI outcomes.

Testicular expression of synaptonemal complex protein 3 (SYCP3) messenger ribonucleic acid in 110 patients with nonobstructive azoospermia.

OBJECTIVE: To determine the expression of the synaptonemal complex protein-3 (SYCP3) gene in men with nonobstructive azoospermia. DESIGN: Cross-sectional case study. SETTING: Avesina Infertility Clinic, Tehran, Iran. PATIENT(S): One hundred and ten consecutive infertile men presenting nonobstructive azoospermia. INTERVENTION(S): Testicular biopsies for histopathological assessment and analyses of SYCP3 expression level by semiquantitative nested reverse transcription-polymerase chain reaction (RT-PCR). The SYCP3 levels were normalized to expression of the housekeeping phosphoglucomutase 1 gene. MAIN OUTCOME MEASURE(S): Expression of SYCP3 messenger ribonucleic acid (mRNA). Correlation of the histopathological findings with SYCP3 expression levels. RESULTS(S): Testicular SYCP3 mRNA expression was observed in 67/110 (60.9%) patients. The expression level correlated with the degree of spermatogenic failure. Although it was expressed in patients with spermatogenesis and maturation arrest, a lack of expression was seen in all of those men with spermatogonial arrest, Sertoli cell-only syndrome, and testicular atrophy. CONCLUSION(S): These data indicate that SYCP3 is expressed in human testis and is restricted to germ cells. Our findings, in association with those obtained in experimental animals, shows that lack of SYCP3 expression in human testis may have a negative effect on spermatogenesis and male fertility.