Normospermic Group Research Articles

Herbicide residues as a possible risk factor in semen quality and spermatogenesis

Background Reports from various countries have consistently demonstrated a correlation between herbicide exposure, arising from agricultural practices, and a decline in semen quality, leading to male infertility. This study was conducted at Ekiti State University Teaching Hospital in Ado-Ekiti, a rural community characterized by a predominantly agrarian population to determine the relationship between semen quality and herbicides residues. Methods The study focused on males whose spouses were seeking assistance at infertility clinics. Routine semen analyses were performed according to the World Health Organization (WHO) criteria, categorizing samples into normospermic, asthenospermic, oligospermic, and azoospermic groups. Seminal plasma samples from each group (twenty samples per group) were subjected to analysis for the presence and concentration of herbicides using High-Performance Liquid Chromatography (HPLC). The following herbicides were investigated: halosulfurum, linuron, fluometuron, chlo-rimuron, imaxamox, cloransulam, dicamba, fluroxypor, trichlopyr, propanil, cloclinafop, clethodim, quizalofop, fluazifop, pinoxaden, bentazon, atrazine, and bromoxynil. The obtained results were subjected to statistical analysis using SPSS version 24. Results The analysis revealed significantly higher concentrations of most herbicides in the asthenospermia, oligospermia, and azoospermia groups compared to the normospermic group (P<0.05). These findings suggest a strong association between herbicide exposure and poor semen quality in the studied population.Conclusion This study provides compelling evidence supporting the hypothesis that herbicides exposure could be a contributory factor to diminished semen quality in the investigated rural com-munity. The results underscore the importance of considering seminal herbicide determination as a routine component in male infertility testing. Additionally, the study advocates for the implementation of relevant legislation to mitigate potential risks associated with herbicide exposure.

Comparison of sperm carnitine profiles of normospermic, oligospermic and azospermic individuals.

The World Health Organization recognizes infertility as a public health issue. An estimated 48.5 million couples worldwide grapple with infertility. Infertility and spermatogenesis dysfunction arise from diverse genetic factors, including single-gene mutations and chromosomal abnormalities. Current research continues to explore other potential causes of infertility, leveraging both proteomic and metabolomic analyses. The primary aim of this study is to underscore the significance of investigating male infertility from a metabolomic perspective. This study aimed to identify new markers for diagnosing and treating male infertility by examining the carnitine profiles in seminal fluids of individuals diagnosed with the normospermic group, oligospermic group, and azospermic group, employing the LC/MS-MS method. The first three L-carnitines C2 (Acetylcarnitine), C8:1 (Octenoylcarnitine), and C16:1 (Palmitoleylcarnitine), emerged as potential novel markers for diagnosis and treatment. Based on these findings, we posit that the results obtained in this study will aid in diagnosing, treating, and monitoring systemic diseases, and provide a foundation for more comprehensive future molecular studies aimed at enhancing prevention and treatment strategies for infertility.

Correlation between sperm parameters and circulating thyroid hormones and testosterone concentrations in Labrador Retriever dog

Thyroid hormones physiologically regulate the semen quality, by altering testosterone concentrations, and different seminal parameters, as well as sperm motility, viability and semen volume. Serum thyroxine (T4), free thyroxine (fT4), thyroid-stimulating hormone (TSH) and testosterone (T) concentrations were measured in 26 adult Labrador Retriever dogs (n. 20 normospermic and n. 6 azoospermic groups), aged 3 ± 0.5 years old, to determine their specific references, by taking into account the related conventional and kinematic sperm parameters and clinical ultrasound evaluations. The mean values of T4, fT4, TSH and T concentrations, as soon as those of sperm parameters and clinical evaluations of normospermic group were in line with dog’s physiological range of literature data. Normospermic group showed significant correlations between T4 and T (r = −0.681; p < .01), sperm progressive motility (%) (r = −0.623; p < .01), and sperm non-progressive motility (%) (r = 0.625; p < .02). The azoospermic group showed higher T4, fT4, TSH and lower T concentrations, compared to normospermic group, and a positive correlation between T4 and T (r = 0.8548; p < .046). The use of breed-specific hormonal ranges and sperm parameters will improve the knowledge of their interaction in Labrador Retriever dogs, adding a new segment of scientific literature. Highlights Thyroxine (T4), free thyroxine (fT4), thyroid stimulating hormone (TSH) and testosterone (T) concentrations were measured in male dogs. Conventional and kinematic sperm parameters were also evaluated. Dogs were divided in 20 normospermic and n. 6 azoospermic Labrador Retrivers. Azoospermic group showed higher values of T4, fT4, TSH and lower values of T compared to normospermic Use of breed-specific hormonal ranges and sperm parameters will improve the knowledge of scientific literature.

Impact of Seminal Plasma Trace Elements on Human Sperm Motility Parameters.

Human seminal plasma contains a variety of macro and trace elements including magnesium (Mg), copper (Cu), zinc (Zn), and iron (Fe) that have essential roles in normal functioning of semen and its quality. The imbalance of these elements has been reported in several pathologic and male infertility disorders. Therefore, this study aimed to determine the levels of these elements in seminal plasma samples, their relationships with each other and their impact on sperm motility. Overall, 192 males (96 normospermic and 96 asthenospermic males) were enrolled in the study. Semen samples were collected by masturbation and computer-assisted/aided semen analysis of sperm motility was performed. The samples were centrifuged and seminal levels of Mg, Cu, Zn and Fe were measured using atomic absorption spectroscopy. The levels of Zn did not differ between the two groups, while the levels of Mg, Cu, and Fe were significantly higher in normospermic males. Fe showed a positive correlation with Mg and Cu in asthenospermic group. However, a negative relationship was found between Mg and Fe levels and between Mg and sperm concentration in the normospermic group. Fe levels were higher in the normospermic group compared to the asthenospermic group. Nevertheless, increased Fe levels caused a decrease in most of sperm motility fractions. Elements play major roles in male fertility and directly affect sperm quality. According to the results of this study, the levels of Zn do not affect the sperm quality and motility, while Fe, Cu and Mg are decreased in males with sperm motility problems. Nevertheless, Fe levels can adversely affect sperm motility in normospermic men.

Association between seminal plasma neopterin and oxidative stress in male infertility: A case-control study

Neopterin is a significant and sensitive marker in estimating the activity of cellular immune system. Oxidative stress plays a role in the etiology of male infertility. Increased reactive oxygen species is accompanied with increase in neopterin level. Hence neopterin may be involved in male infertility. The objective of this case-control study was to determine neopterin level in idiopathic infertile and normospermic men; furthermore, to identify its relationship with oxidative stress markers including total oxidant, malondialdehyde, sperm DNA fragmentation, and total antioxidant capacity of seminal plasma. Forty seven infertile and forty three normospermic males were selected according to WHO criteria. Their semen and blood samples were taken; subsequently, the levels of neopterin, total oxidant, total antioxidant, malondialdehyde, and sperm DNA fragmentation were measured. The levels of neopterin, total oxidant, and malondialdehyde in seminal plasma of infertile males were significantly higher than those of normospermic group (p=0.038, 0.018, and 0.028, respectively). Furthermore, sperm DNA fragmentation in infertile men was higher than that of control group (p<0.001). Moreover, total antioxidant capacity of seminal plasma in infertile males was significantly lower than that of normospermic subjects (p=0.002). No significant difference was observed in serum neopterin, total oxidant, and malondialdehyde between the infertile and normospermic groups. The significant inverse correlation between seminal plasma neopterin and total antioxidant in the infertile males supports a possible role of neopterin in male infertility. Neopterin can be suggested as a marker in monitoring and diagnosis of idiopathic male infertility.

Examination of Y-Chromosomal Microdeletions and Partial Microdeletions in Idiopathic Infertility in East Hungarian Patients.

Purpose:The aim of this study was to establish the Y chromosome microdeletion and partial AZFc microdeletion/duplication frequency firstly in East Hungarian population and to gain information about the molecular mechanism of the heterogeneous phenotype identified in males bearing partial AZFc deletions and duplications.Materials and Methods:Exactly determined sequences of azoospermia factor (AZF) region were amplified. Lack of amplification was detected for deletion. To determine the copy number of DAZ and CDY1 genes, we performed a quantitative analysis. The primers flank an insertion/deletion difference, which permitted the polymerase chain reaction products to be separated by polyacrylamide gel electrophoresis.Statistical Analysis Used:Mann–Whitney/Wilcoxon two-sample test, Kruskal–Wallis test, and two-sample t-probe were used for statistical analysis.Results:AZFbc deletion was detected only in the azoospermic cases; AZFc deletion occurred significantly more frequently among azoospermic patients, than among oligozoospermic males. The frequency of gr/gr deletions was significantly higher in the oligozoospermic patients than in the normospermic group. The b2/b3 deletion and partial duplications were not different among our groups, while b1/b3 deletion was found only in the azoospermic group. In infertile males and in normozoospermic controls, similar Y haplogroup distribution was detected with the highest frequency of haplogroup P. The gr/gr deletion with P haplogroup was more frequent in the oligozoospermic group than in the normozoospermic males. The b2/b3 deletion with E haplogroup was the most frequent, found only in the normozoospermic group.Conclusions:Y microdeletion screening has prognostic value and can affect the clinical therapy. In case of Y chromosome molecular genetic aberrations, genetic counseling makes sense also for other males in the family because these types of aberrations are transmittable (from father to son 100% transmission).

Cryopreservation triggers DNA fragmentation and ultrastructural damage in spermatozoa of oligoasthenoteratozoospermic men

Objective: Oligoasthenoteratozoospermia (OAT) is one of the causes of male infertility. Cryopreservation is valuable for the management of infertility. This study aimed to reveal the effects of cryopreservation on sperm motility, morphology, ultrastructural details and DNA fragmentation in patients with OAT. Materials and Methods: In this observational study, ejaculates were collected from 40 male volunteers of whom 20 were OAT and 20 were normospermic. The ejaculates were stored in liquid nitrogen at -196oC and analysed following thawing after 1 or 3 months. Ejaculates were evaluated in terms of motility, morphology and DNA fragmentation before and after thawing. Results: Sperm motility and morphology were both affected by cryopreservation in samples from both groups. After thawing, spermatozoa with morphological abnormalities were increased significantly in both groups. Ultrastructural investigations showed alteration in integrity of the membranes and increased acrosomal defects. Post-thaw investigations revealed prominent increases in the number of DNA fragmented cells in both groups when compared to the results before freezing. OAT groups revealed a significantly higher number of DNA fragmented spermatozoa compared to the normospermic group for both time periods. Conclusion: Cryopreservation produces ultrastructural damage and induces DNA fragmentation in both normospermic and OAT groups. The damage is more severe in the OAT group.

MP35-10 RELATIONSHIP BETWEEN SEMINAL PARAMETERS AND SPERMAL DNA FRAGMENTATION

You have accessJournal of UrologyInfertility: Epidemiology & Evaluation II1 Apr 2017MP35-10 RELATIONSHIP BETWEEN SEMINAL PARAMETERS AND SPERMAL DNA FRAGMENTATION Jose Olmedo, Rosa Molina, Andrea Tissera, Enzo Malizia, and Tristan Dellavedova Jose OlmedoJose Olmedo More articles by this author , Rosa MolinaRosa Molina More articles by this author , Andrea TisseraAndrea Tissera More articles by this author , Enzo MaliziaEnzo Malizia More articles by this author , and Tristan DellavedovaTristan Dellavedova More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.1100AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Over the years, a large number of tests have been proposed for clinical evaluation of semen quality, some are used in clinical practice, but few are routinely used. Given the complexity of the sperm cell, there is no single assay capable of providing complete information for diagnosis and prognosis of fertility. Objective: To evaluate the correlation between DNA fragmentation and seminal parameters in patients who consulted for fertility. Design: Observational cross-sectional retrospective. METHODS Materials and Methods: 1,562 samples man who consulted for fertility were studied. DNA fragmentation was performed by the TUNEL technique. (In Situ Cell Death Detection Kit Roche). A percentage of fragmentation <20% was used as the normal reference value. Seminal parameters were analyzed according to WHO Standards 2010. Statistic: T-test was used for independent samples. The Chi2 ratios and the correlation of TUNEL with respect to seminal parameters were analyzed with Spearman's test. A p <0.05 was considered statistically significant. RESULTS Results: In the total group, 31.6% (494/1562) presented increased fragmentation. The proportion of men with increased TUNEL, according to the different seminal parameters was: Normal concentration (29%, 381/1319) vs Oligozoospermia (47%, 113/243); P <0.0001. Normal Mobility (25%; 291/1157) vs. Astenozoospermia (60%); P <0.0001. Normal morphology (24%) vs Teratozoospermia (50%, 203/405); P <0.0001. Patients with oligoastenoteretozoospermy (OAT) had a higher proportion of TUNEL increased, 53.7% [58/108], compared to the Normospermic group 22.4% (173/772), P <0.0001. We found no difference in the fragmentation between groups of patients with Toxic habits (Alcohol and Tobacco) and without them. Of patients exposed to agrochemicals, 45.3% showed increased DNA fragmentation. A significant and negative correlation was found between TUNEL and concentration / ml, total concentration of the ejaculate; total motility; rapid progressive motility; morphology and nuclear maturity. (P <0.0001). CONCLUSIONS Conclusion: According to our results, we observed a significant negative correlation between TUNEL and seminal parameters. In agreement with other works we find that the exposure to environmental agrochemicals would produce a deleterious effect on the fragmentation of the DNA. Taking into account that an increased TUNEL was found in 22.4% of Normospermic patients, we emphasized the usefulness of this test to more fully study the fertility of the patients and thus to be able to take therapeutic measures that favor reproductive success. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e461 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Jose Olmedo More articles by this author Rosa Molina More articles by this author Andrea Tissera More articles by this author Enzo Malizia More articles by this author Tristan Dellavedova More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Effect of gr/gr microdeletion in partial AZFc deletion on human spermatogenesis

Objectives To inverstigate the effect of gr/gr subdeletion in partial AZFc deletion on human spermatogenesis. Methods We assessed The prevalence of gr/gr deletion in a population of both normospermic and infertile men with severe impairment of spermatogenesis were assessed, the contribution of these gr/gr deletion to male infertility was determined. A combined methodological approach that included the detection of the different DAZ gene copies by sequence nucleotide variant (SNV) analysis and AZFc-specific sequence tagged site (STS) markers was used. Results Type of gr/gr deletion was detected in 27 cases of 278 males with oligozoospermic and 17 cases of 220 donors. There was no difference in the prevalence of gr/gr deletion comparing oligozoospermic and normospermic groups (P=0.438). Of the 27 men with the gr/gr deletion, 23 cases exhibited the doublet deletion of DAZ1/DAZ2 and 1 case showed the DAZ3/DAZ4 deletion. In the normozoospermic population, 16 cases of gr/gr-DAZ3/DAZ4 deletions and one gr/gr-DAZ1/DAZ2 deletion were found. There was a significant difference when comparing gr/gr-DAZ1/DAZ2 deletions vs. gr/gr-DAZ3/DAZ4 deletions in the oligozoospermic and normospermic groups (P=0.000). Conclusions The gr/gr-DAZ3/DAZ4 deletion may not be sufficient to cause spermatogenic impairment. In addition, gr/gr microdeletions, which involve the DAZ1/DAZ2 region, may be a main cause of low sperm concentration and spermatogenic failure in the south chinese population. Key words: Spermatogenesis; Y Chromosome

Detection of subpopulations of leucocytes in different subgroups of semen sample qualities

The role of leucocytospermia in male subfertility is a much debated topic despite being a frequent finding. This study aimed to identify the role of leucocytes, leucocyte subpopulations and natural killer cells in male subfertility. Seventy-sex subfertile men attending a regional andrology unit were recruited into this prospective study and subdivided into groups based on their semen analysis. The different leucocyte subpopulations were identified using immunocytochemical staining. Significant levels of CD3 helper T lymphocytes (P < 0.001) were present in the oligospermic, asthenospermic, oligoasthenospermic and obstructive azoospermic group compared to the normospermic group. Significant levels of B cells (P < 0.05) were present in the asthenospermic, oligoasthenospermic and obstructive azoospermic group. The natural killer cells (CD56) were significantly raised in the oligoasthenospermic and obstructive azoospermic group (P < 0.05). Our study suggests that leucocytospermia impairs sperm function through enhanced T helper cell modulation, increased B cell population which leads to increased levels of antisperm antibody and natural killer cells mediated sperm damage. The site of seminal leucocyte production is not necessarily confined to the vas or the epididymis.

Assessment of Seminal Estradiol and Testosterone Levels as Predictors of Human Spermatogenesis

The proposed hypothesis for this study was that seminal testosterone/estradiol levels and/or their ratios may be a good indicator for predicting normal spermatogenesis. The concentrations of estradiol and testosterone in seminal fluid were measured using competitive immunoassay techniques in specimens collected from 192 infertile patients and 103 normospermic men. Infertile patients were subdivided into three groups according to their semen analysis results and testicular biopsy: oligozoospermia, obstructive azoospermia (OA), and nonobstructive azoospermia (NOA). Results showed that seminal testosterone levels in the infertile groups were lower than in the normospermic individuals (P < .01), whereas seminal estradiol levels in the OA group were significantly higher than those in normospermic and NOA groups (P < .01). Testosterone/estradiol ratios in the seminal plasma from the infertile groups were significantly lower than that in the normospermic group (P < .01). However, seminal estradiol levels among normospermic and NOA groups showed no significant differences. These results suggest that the local balance between androgen and estrogen, or their ratios, may play an important role in maintaining normal spermatogenesis. Also, decreased seminal testosterone/estradiol ratio may be a good indicator for identifying the absence of sperm production in NOA patients.

Enhancement in motility of sperm co-incubated with cumulus oocyte complex (COC) in vitro

Objectives To study the influence of cumulus cell co-culture on sperm motility under in vitro conditions. Study design Patients recruited for IVF were included in this prospective study. Among 30 subjects, 15 were normozoospermic and 15 were teratozoospermic. The oocyte cumulus mass was inseminated with optimum number of spermatozoa. Sperm motility was documented every 24 h after insemination. Results Sperm motility without co-culture declined with incubation time and complete loss of motility was observed at the end of 72 h. The inclusion of cumulus cells with sperm almost doubled the sperm motility at 48 h incubation time. At 72 h incubation time, the sperm + cumulus group had 14% total motility in comparison to 0% in the sperm alone group. When compared to the teratospermic group, the normospermic group did not show any significant difference with respect to sperm motility at 24 and 48 h both in the sperm alone and the sperm + cumulus groups. However at 72 h significantly higher ( P < 0.05) motility was maintained in the normospermic group. Conclusion The result of the present investigation suggests that co-culture with cumulus cells enhances sperm motility under in vitro conditions.

Phenotypic variation within European carriers of the Y-chromosomal gr/gr deletion is independent of Y-chromosomal background

Background:Previous studies have compared sperm phenotypes between men with partial deletions within the AZFc region of the Y chromosome and non-carriers, with variable results. In this study, a separate question...

Seminal Plasma Zinc Concentration and α-Glucosidase Activity with Respect to Semen Quality

This study was conducted to examine the relationship between the concentration of zinc and neutral alpha-glucosidase (NAG) with semen quality. Semen samples from 75 male partners of couples who were attending the Obstetrics and Gynecology Department were analyzed for semen quality. Based on sperm count, the subjects were divided into three groups. Zinc and neutral alpha-glucosidase activity were estimated in seminal plasma. Results showed that mean the alpha-glucosidase activity was lowest among the azoospermic group with respect to oligozoospermic and normozoospermic groups. Mean zinc levels were also lower among azoospermics compared to oligozoospermic and normospermic groups. The difference was statistically significant (p < 0.05). A significant positive correlation was observed between zinc levels and sperm count (r = 0.29, p < 0.05) and zinc and alpha-glucosidase activity (r = 0.31, p < 0.05) in seminal plasma. These results suggest that zinc and neutral alpha-glucosidase seem to play an important role in human reproduction.

SCREENING OF Y CHROMOSOME MICRODELETIONS IN TUNISIAN INFERTILE MEN

The aim of this study was to establish the prevalence of Y chromosomal microdeletions in infertile Tunisian men. Three groups of infertile men, 65 normospermic, 53 oligozoospermic and 45 azoospermic, were tested for Yq microdeletions detection by multiplex polymerase chain reaction (PCR) using specific Y chromosome AZF regions tagged site markers (STS). One group of 13 healthy men was used as the control group. Six STS were tested (2 in each AZF region). The general prevalence of AZF microdeletions was 16%; in azoospermia and severe oligospermia groups, it was higher (29% and 30.5%, respectively). Significant differences were found with moderate oligospermic and normospermic groups (p < 0,05). AZFc microdeletions were the most frequent, and 55% of AZFc deleted patients were oligospermic. No deletions were detected in the control group. These results add to the growing literature data, showing that microdeletions of the Y chromosome is an important cause of severe spermatogenetic defect and confirm that deletion in AZFc region is the most common and is compatible with residual spermatogenesis.

GR/GR deletions within the azoospermia factor c region on the Y chromosome might not be associated with spermatogenic failure

No statistically significant difference in deletion frequency was found between infertile patients and a control group. It is suggested that in some populations the gr/gr deletion might be an inconsequential polymorphism.

Relationship of serum and seminal plasma zinc levels and serum testosterone in oligospermic and azoospermic infertile men.

To measure the zinc level in fertile and infertile population and to find out relationship of serum and seminal plasma zinc levels and serum testosterone in oligospermic and azoospermic infertile men. Comparative study. The Infertility Clinic, Jinnah Postgraduate Medical Center (JPMC), Karachi, from January 2002-July 2003. Fifty-eight primary infertile male subjects, without any treatment, who had regular unprotected intercourse for at least 12 months without conception with their partners, aged 20-40 years, having normal and regular menstrual cycles without any uterine pathology or hormonal disturbance were selected from Infertile Clinic, Jinnah Postgraduate Medical Center, Karachi. After semen analyses, they were grouped as, oligospermic (30), and azoospermic (28). Twentyfive known fertile male selected from general population (their spouse had given birth to a child within one year) and after semen analysis were taken as normospermic control group . Semen analyzed according to WHO criteria and serum and seminal plasma zinc was estimated by 5Br. PAPS colorimetric method. Serum testosterone, LH, and FSH were measured by ELISA, using kits supplied by Diagnostic System Laboratories (DSL), U.S.A. The mean value for testosterone was significantly low in oligospermic, (8.04 -/+ 0.76) and azoospermic subjects (7.14 -/+ 0.27) as compared to normospermic (9.90 -/+ 0.14) control groups (p<0.05, p<0.001 respectively), serum LH (p<0.05, p<0.001 respectively) and FSH (p<0.05) were significantly higher than normospermic control group. Serum and seminal plasma zinc levels was low in oligospermic, and azoospermic subjects when compared with normospermic control groups (p<0.05, p<0.001 respectively). This suggests its involvement in spermatogenesis and infertility in such patients. Seminal zinc level has weak correlation with testosterone in oligospermic (r=0.44, p<0.05) and with serum zinc in azoospermic subjects (r=0.49, p<0.05) while no correlation was observed with LH and FSH in all the groups. In this study, there was significant decrease in serum and seminal plasma zinc levels in oligospermic and azoospermic infertile males with significantly low androgen. It indicates zinc has a possible role for spermatogenesis and steroidogenesis. Therefore, zinc concentration in seminal plasma should be considered as one of the factors responsible for decreased testicular function in infertile male subjects.

Frequency of disordered zona pellucida (ZP)-induced acrosome reaction in infertile men with normal semen analysis and normal spermatozoa-ZP binding.

Results of zona pellucida (ZP)-induced acrosome reaction (AR) are reported for 186 normospermic men with unexplained infertility and compared with 34 normal fertile men and 54 patients with disordered ZP-induced AR (DZPIAR) diagnosed after failure of standard IVF. For each ZP-induced AR test, four oocytes that failed to fertilize in IVF were incubated for 2 h with 2x10(6)/ml motile spermatozoa. Spermatozoa tightly bound to the ZP were recovered by aspirating the oocytes with a pipette and the AR assessed using pisum sativum agglutinin labelled with fluorescein. The standard deviation of the difference was 5.2% for repeated tests for ZP-induced AR on different ejaculates from 54 men. The ranges for the ZP-induced AR were 3-98% for normospermic infertile men, 24-95% for fertile men and 0-16% for DZPIAR patients. In the normospermic group, there was a significant correlation between ZP-induced AR and sperm concentration (Spearman r = 0.238, P < 0.001). Using ZP-induced AR < or =16% as the threshold for diagnosis of DZPIAR, the frequency of this condition in normospermic infertile men would be 25%. Thus DZPIAR is common with normospermic idiopathic infertility and this condition should be diagnosed before assisted reproductive technology since it requires intracytoplasmic sperm injection.

Biochemical markers of early and late spermatogenesis: relationship between the lactate dehydrogenase-X and creatine kinase-M isoform concentrations in human spermatozoa.

As part of our research program on biochemical markers of sperm maturity, we have studied sperm creatine kinase (CK) and lactate dehydrogenase (LDH) concentrations and the isoform ratios of the CK-M [% CK-M/(CK-M + CK-B)] and LDH-X [% LDH-X/(LDH-X + LDH-a)] in 50 oligospermic and 95 normospermic men [corrected]. Because the synthesis of LDH-X is initiated in early spermatogenesis, and that of CK-M commences in late spermiogenesis simultaneously with cytoplasmic extrusion, we proposed two working hypotheses:(1) LDH and CK concentrations reflect the retained cytoplasm in sperm, thus the activities of both enzymes will be related and will be higher in oligospermic specimens, which have a higher incidence of immature spermatozoa; and (2) because in normally developed sperm both LDH-X and CK-M are present, there will be a correlation between LDH-X and CK-M ratios in the mature sperm populations. However, among men with immature sperm samples with late spermiogenetic defect and diminished CK-M ratios, there will be two groups: one which completed spermatogenesis prior to spermiogenetic failure (normal LDH-X and diminished CK-M ratios), and another group with defects in both spermatogenesis and spermiogenesis (low LDH-X and diminished CK-M ratios). Because of this heterogeneity, LDH-X ratios will be a poor predictor of sperm maturity. The data support the hypotheses: (1) LDH and CK concentrations were higher in oligospermic vs. normospermic men (P < 0.001). (2) The LDH and CK concentrations were related (r = 0.65, P < 0.001, N = 145), and there were inverse correlations between CK, LDH, LDH-X, or CK-M ratios vs. sperm concentrations (P < 0.001 in all four). (3) The CK-M and LDH-X ratios were different between the oligospermic and normospermic groups (P < 0.001), although the means of the LDH-X ratios were narrower (LDH-X:1:1.3; CK-M:1:1.9). (4) Dividing the 145 samples by the cut-off value of mean minus 1 SD of the CK-M and LDH-X ratios (11% and 32%, respectively) demonstrated that the CK-M ratios discriminated better than LDH-X ratios between the samples with mature and immature sperm. These data on the biochemical markers of early and late spermatogenesis support the studies in which CK better reflected sperm quality than LDH or LDH-X (Orlando et al., 1994: Int J Androl 17:13-18) and the > 10% sperm CK-M ratio predicted with a rate of 30.4% per cycle in the occurrence of pregnancies in a blinded study of 84 IVF couples (Huszar et al., 1992: Fertil Steril 57:882-888).

How often should infertile men have intercourse to achieve conception?

To clarify how often infertile men should have intercourse to achieve conception, the effect of sequential ejaculation on total motile sperm counts was investigated. Case-control study. Infertility and IVF unit, tertiary care center. Five hundred seventy-six men who produced two closely spaced sequential ejaculates. The total motile sperm counts of the second ejaculates were compared with the total motile sperm counts of the first ejaculates. In normospermic men (n = 359), the total motile sperm counts decreased significantly from 93 (18 to 601) (median [minimum to maximum] x 10(6)) in the first ejaculate to 42 (1.2 to 387) in the second ejaculate, produced 24 hours later. In contrast to the normospermic men, in the asthenospermic group (24 hours difference, n = 81) and in both oligospermic groups, (1 to 4 hours difference, n = 27; and 24 hours difference, n = 45), there were no significant changes in the total motile sperm counts (24 [5.9 to 229] versus 30 [0.8 to 150], 6 [0.8 to 18] versus 3.6 [0.1 to 63] and 13 [2.5 to 32] versus 10 [0.1 to 66], respectively). Moreover, in both oligoasthenospermic groups (1 to 4 hours difference, n = 23; and 24 hours difference, n = 41) the total motile sperm counts increased significantly (3.2 [0.6 to 7.9] versus 8 [0.4 to 48] and 4 [0.2 to 13] versus 4 [0.1 to 101], respectively). In all groups, pooling sequential ejaculates significantly increased the total motile sperm counts, over and above that of the first ejaculate, by 49% in the normospermic group, 95% in the asthenospermic group, 67% and 75% in the oligospermic groups (1 to 4 hours and 24 hours difference, respectively), and 233% and 139% in the oligoasthenospermic groups (1 to 4 hours and 24 hours difference, respectively). Sequential ejaculation may overcome the impaired sperm transport causing low total motile sperm counts observed in some oligospermic and/or asthenospermic men. Most of these infertile men may significantly increase their fertility potential, assessed by the total motile sperm counts, either by pooling sequential ejaculates for IUI, GIFT, and IVF, or by having intercourse every day or even twice a day, at the time of ovulation.