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The objective of the present study was to evaluate the association between the expression of sperm mannose-ligand receptors and sperm morphology. Sperm samples were obtained from 45 men, 30 fertile sperm donors and 15 infertile men. Sperm concentration, motility and morphology were evaluated and then incubated with control medium (Ham's F-10 + 1% HSA) for 4 h. Expression of mannose-ligand receptors was evaluated by mannosylated-BSA-FITC (subdivided into 3 patterns: I, for uncapacitated sperm; II, for capacitated; and III, for acrosome-reacted sperm). The mean ( ±SE) frequencies of sperm cells of the total sperm population that expressed patterns I, II, and III were 88 ±2.1%, 7 ±1.6%, and 5 ±0.8%, respectively, for fertile men, and 90 ±2.1%, 7 ±1.3%, and 3 ±0.5%, respectively, for infertile men. The rate of pattern III expression of mannose-ligand receptors was significantly higher in the fertile group compared to the infertile patients ( p < .01). A poor but significant correlation was observed between the rate of pattern III and the percentage of normal-forms sperm cell in the ejaculate ( r = .35, p = .018). Fertile sperm samples express more advanced patterns of mannose-ligand receptors compared to infertile men. This phenomenon is related to the morphology of human sperm cell in the ejaculate more than to any other basic sperm characteristics.

The study was conducted to evaluate the significance of preoperative clinical parameters for detection of mature testicular sperm cells in nonobstructive azoospermic men. Sixty-five consecutive men with nonobstructive azoospermia underwent testicular sperm extraction procedures. Testicular samples were analyzed histologically with patterns classified as mature spermatogenesis (normal or partial), arrest of spermatogenesis, and Sertoli cell only. Testicular sperm cells were isolated for use in an IVF/ICSI program. Histologic patterns and detection rate of sperm cells were correlated to clinical characteristics. Mature sperm cells were found in all levels of serum FSH. The men were divided into 3 groups based on their clinical characteristics (serum FSH level and testicular size). The distribution of the different testicular histologic patterns, as well as detection rate of sperm cells, was similar in all groups. No correlation was found between serum levels of FSH, LH, prolactin, or testosterone and sperm presence. None of these parameters, nor the testicular size and consistency, can serve as predictive variables of the histological pattern or the presence of mature sperm cells in the testicular biopsies in cases of nonobstructive azoospermia. Until an effective predictive tool is available, a trial of sperm retrieval is recommended for all azoospermic men independent of their clinical characteristics.

Objective: To investigate placental apoptosis in discordant dichorial twins. Methods: Placental samples were obtained from 7 third-trimester suitable twins. Discordancy was defined as a >25 per cent difference in newborn birth weight. Light microscopy using hematoxylin and eosin (H&E)-stained paraffin slides and terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end-labelling (TUNEL) methods were used to confirm the incidence of apoptosis. Investigators were blinded to pregnancy outcome. Results: Both methods revealed that the incidence of apoptosis in the placentas of the smaller fetuses was significantly higher than in placentas of the larger fetuses. The incidence of TUNEL-positive cells in the former was 1.4±0.26 per cent: this was significantly higher than the incidence of apoptosis in the placental specimens of the latter (0.9±0.07 per cent, P&lt; 0.02 Wilcoxon rank test). The same results were obtained with H&E: the incidence of apoptosis detected in placentas from the former was 1.07±0.1 per cent compared to 0.72±0.08 per cent in those of the latter (P&lt; 0.02 Wilcoxon rank test). Conclusions: Despite similar environment conditions, placental apoptosis is increased in the smaller fetus and thus might play a role in discordancy between twins. Since increased placental apoptosis has also been found in singleton intrauterine growth restriction, this supports the hypothesis that the smaller twin is selectively growth restricted. Copyright 2002 Elsevier Science Ltd

BACKGROUND: Artificial insemination by donor spermatozoa (AID) can prove a valuable treatment for a number of male factor disorders, although its success rate is variable. METHODS: Retrospective analysis of the results of 6139 cycles performed in 1001 women during an 18 year period is presented. Pregnancy rates per cycle are presented as a function of: female fertility history, treatment modalities, medication used for induction of ovulation, female age, year of treatment, consecutive cycle effect and the use of fresh versus frozen–thawed spermatozoa. RESULTS: Overall pregnancy rate of 12.6% and cumulative pregnancy rate after 12 months of treatment of 75% were achieved. Age was found to be the most important determinant for success rate. CONCLUSIONS: Since the establishment of AID treatments, the mean age of the population of women receiving treatment has increased each year. Consequently, success rate did not improve, even with the use of more sophisticated medical modalities.

Artificial insemination by donor spermatozoa (AID) can prove a valuable treatment for a number of male factor disorders, although its success rate is variable.

The objective of the present experiments was to establish an acceptable standard for the cervical mucus penetration test (CMPT) by determining the minimal progressive motile spermatozoa concentration (PMSC) that will yield the highest score for proven fertile donor sperm specimens. For this purpose, fresh and frozen-thawed samples were used. Semen was obtained from 29 fertile donors and different PMSC (8, 10, and 14 106/mL) were prepared for each sample. The same mucus specimen was used for testing each sperm sample in the three different dilutions. No difference in the scoring of the CMPT between the fresh vs. frozen-thawed groups was found. When PMSC of 14 106/mL was used, almost all specimens scored the highest rank. The present study revealed that only semen samples with a minimal PMSC of 14 106/mL cells can be used in the CMPT. The information that the freeze-thaw process does not affect the CMPT results supports the concept of cryopreservation of pooled fertile donor specimens in aliquots with adequate concentration of progressive motile spermatozoa for later use as a CMPT control.

Recently, microdeletions in the azoospermic factor region of the Y chromosome, in addition to chromosomal anomalies, have been detected in men with azoospermia or severe oligozoospermia. In this study we evaluated the molecular and cytogenetic defects of infertile men. The frequency of Y microdeletions among 105 azoospermic, 28 oligozoospermic and 32 fertile men was tested on lymphocyte DNA using a series of 20 sequence-tagged sites. In addition, microdeletions were evaluated on testicular-derived DNA among 26 azoospermic patients who underwent testicular biopsy and in whom no sperm cells could be identified. Karyotype analysis was performed on 72 of the infertile patients. Deletions were detected in 6.7% azoospermic and 3.6% oligozoospermic men. No deletions were identified among the fertile men. Identical results were obtained with DNA derived either from lymphocytes or testicular tissue. The frequency of chromosomal aberrations in the 72 infertile patients tested (62 azoospermic, 10 oligozoospermic) was 16.6%, with a high percentage of gonosome anomalies. Additional andrological parameters (hormone values, cryptorchidism) failed to identify men at risk for having microdeletions before the test. Our findings support the recommendation to perform genetic defect screening among infertile men before their enrollment in an intracytoplasmic injection/in-vitro fertilization programme.

The present study was conducted to assess the fertilizing potential of frozen-thawed spermatozoa, which were cryopreserved after separation on a Percoll gradient, or washed out of seminal plasma. For this purpose, binding to the zona pellucida and other characteristics of the treated sperm cells were compared with those of cryopreserved spermatozoa from the same original sample which were not manipulated before freezing. Semen specimens were obtained from 80 candidates for sperm donation. Percoll-treated sperm samples compared with the sibling, unprocessed controls had significantly higher values of sperm motility characteristics and per cent of cells with normal morphology after freezing and thawing. Sperm binding ability to the zona pellucida was not statistically different (109 +/- 8.1% and 94 +/- 6.7% in unprocessed and Percoll-treated samples respectively). Sperm specimens processed by washing had significantly higher values for motility characteristics than untreated sibling samples, but no differences were found between the treated and untreated samples for morphology and binding to the zona pellucida (hemizona index of 75 +/- 7.0% and 76 +/- 6.7% in unprocessed and washed samples respectively). These findings suggest that, judged by the binding assay, the aforementioned pre-freezing separation processes have no adverse effect upon the fertilizing potential of the thawed sperm cells. These procedures make it possible to optimize the progressive motile sperm cell concentration of the frozen specimen, which facilitates the storage of samples with good quality, even when the features of the original semen are sub-optimal.

The objective of the study was to evaluate the benefit of different sperm preparation methods by using the hemizona assay. A total of 58 men admitted to the male infertility clinic for evaluation were tested by routine semen analysis and hemizona assay. Five different techniques (swim-up, TEST-yolk buffer, Percoll, pentoxifylline and progesterone) were used for preparation of sperm suspensions. The effect of these treatments on the sperm-binding capacity using the hemizona assay was assessed. The routine swim-up preparation was used as the reference method. Of the four preparation methods, only the TEST-yolk buffer and pentoxifylline exhibited an overall statistically significant improvement in sperm-binding capacity in comparison with the swim-up preparation method (P = 0.01 and 0.001 respectively). Following preparation with Percoll and progesterone there was no change in the mean value of binding capacity, compared with swim-up. However, examination of the effect of the four treatments on each specimen individually yielded a diversity in the response, e.g. having the capability to enhance, damage or be ineffective in sperm binding capacity. The results support the conclusion that in-vitro sperm preparation methods can affect sperm binding to the zona pellucida. Since there is a diversity in the response of sperm samples to different treatments, the hemizona assay can be used in selecting the optimal sperm preparation method prior to its use for assisted reproductive techniques. This is advocated mainly for the 'male factor' group.

We retrospectively analysed the characteristics of 54 men with testicular cancer. The group comprised 32 men with pure seminoma and 22 with non-seminoma germ cell tumours (NSGCT). This group was further compared to 190 healthy sperm donor candidates. Sperm quality was found to be higher in the seminoma versus the NSGCT patients in: sperm concentration [50 (0-230) versus 17 (0-288) x 10(6)/ml, P < 0.001], total motile sperm counts (TMC) [57 (0-508) versus 12 (0-854) x 10(6)/ejaculate, P = 0.002], post-thaw forward motile concentration [3 (0-28) versus 1.7 (0-17) x 10(6)/ml, P = 0.003] and motility percentage [20 (0-57) versus 12.5 (0-42) %, P = 0.002]. Serum hormone concentrations did not differ between these two sub-groups, although the follicle stimulating hormone concentrations were higher than normal in both (14.6 +/- 2.5 versus 10.4 +/- 1.4 mIU/ml, P > 0.05). As is well documented, cancer patients were found to have lower sperm quality compared to healthy candidates. The existence of these differences, and the fact that testicular cancer affects spermatogenesis, indicated that the mechanisms involved in the deterioration of sperm quality can, at least partially, be attributed to the type and origin of the malignant cancer. The higher sperm counts in the seminoma group may be related to the fact that the resemblance of the seminoma cells to normal germ cells is greater than that of the NSGCT cells, and therefore they retain a better capacity to function. Due to modern assisted reproductive technologies and micromanipulation achievements, the lower yield of spermatozoa in severe cases is no longer a major obstacle to offering cryopreservation to these patients.

The study was conducted to determine the deleterious effect of lymphoma disease on spermatogenesis and to evaluate the possibility that the disease is mediated primarily by inherent mechanisms in Hodgkin's disease and non-Hodgkin's lymphoma patients. A total of 89 patients with lymphoma disease (Hodgkin's and non-Hodgkin's) were referred for sperm preservation prior to adjuvant treatments. A comparison was made of pre- and post-thaw sperm quality between lymphoma patients and healthy volunteers who applied for sperm donation. This was followed by further assessment of the differences between patients with Hodgkin's disease and non-Hodgkin's lymphoma in terms of sperm variables, clinical parameters and blood hormone concentrations. It was found that patients with lymphoma disease had significantly impaired pre-freeze and post-thaw sperm quality compared with that of healthy volunteers. Patients with non-Hodgkin's lymphoma had spermatozoa of higher quality than patients with Hodgkin's disease. No differences were found in the clinical or hormonal parameters between these two groups. As expected, reduced testicular size and abnormal testicular consistency were correlated with decreased sperm quality. The mere presence of cancer disease has a direct negative effect on spermatogenesis, which is probably not related to incidental side-effects. A variable degree of impairment should be expected with different categories of cancer.

Spermatogenesis may be focal in non-obstructive azoospermia. The present study was conducted to determine whether the performance of multiple, rather than a single testicular sample contributes to obtaining spermatozoa in amounts sufficient for fertilization and cryopreservation in non-obstructive, azoospermic patients. Furthermore, the aim was to clarify the significance of location for retrieval from the testis in such cases. Three biopsies were taken from identical locations in 55 testes of 29 men with non-obstructive azoospermia: (i) the rete testis region, ii) the midline, and (iii) the proximal region of the testis. When sperm cells were detected, they were used for intracytoplasmic sperm injection (ICSI), and the remainder were then cryopreserved in as many aliquots as possible (adjusted for ICSI procedure). Spermatozoa were found in 28 testes (50.9%) of 18 men (62.1%). In the testes from which spermatozoa were obtained, they were present in three, two or one locations in 15 (53.6%), five (17.9%) and eight (28.6%) cases respectively. The possibility of finding spermatozoa was not influenced by the location in the testis. Multiple testicular sperm extraction is recommended in cases of non-obstructive azoospermia, since it may enhance diagnostic accuracy of absolute testicular failure and increase the number of sperm cells retrieved.

We aimed to evaluate the effect of the levonorgestrel-releasing intrauterine system (LNG-IUS) on the uterine vasculature and the endometrium.

The purpose of this study was to evaluate the effect of female age on the capacity of the zona pellucida to bind spermatozoa. A total of 1008 unfertilized oocytes obtained from 210 women (aged 21-43 years) participating in the in-vitro fertilization programme were tested using a hemizona assay. Spermatozoa taken from a cryopreserved pool of fertile donor specimens served as a control in the hemizona assay, and were used to assess the ability of the zona pellucida to bind spermatozoa. The mean +/- SD number of spermatozoa attached to the hemizona was 107 +/- 42. The binding capacity of different oocytes from the same cohort varied substantially (coefficient of variation = 28%). Age was not found to be correlated with the number of spermatozoa bound to the zona pellucida (r = -0.02; P > 0.1). It was concluded that female age has no role in the ability of the human zona pellucida to bind spermatozoa.

The objective of the study was to evaluate the benefit of different sperm preparation methods by using the hemizona assay. A total of 58 men admitted to the male infertility clinic for evaluation were tested by routine semen analysis and hemizona assay. Five different techniques (swim-up, TEST-yolk buffer, Percoll, pentoxifylline and progesterone) were used for preparation of sperm suspensions. The effect of these treatments on the sperm-binding capacity using the hemizona assay was assessed. The routine swim-up preparation was used as the reference method. Of the four preparation methods, only the TEST-yolk buffer and pentoxifylline exhibited an overall statistically significant improvement in sperm-binding capacity in comparison with the swim-up preparation method (P = 0.01 and 0.001 respectively). Following preparation with Percoll and progesterone there was no change in the mean value of binding capacity, compared with swim-up. However, examination of the effect of the four treatments on each specimen individually yielded a diversity in the response, e.g. having the capability to enhance, damage or be ineffective in sperm binding capacity. The results support the conclusion that in-vitro sperm preparation methods can affect sperm binding to the zona pellucida. Since there is a diversity in the response of sperm samples to different treatments, the hemizona assay can be used in selecting the optimal sperm preparation method prior to its use for assisted reproductive techniques. This is advocated mainly for the ‘male factor’ group.

The purpose of this study was to evaluate the effect of female age on the capacity of the zona pellucida to bind spermatozoa. A total of 1008 unfertilized oocytes obtained from 210 women (aged 21–43 years) participating in the in-vitro fertilization programme were tested using a hemizona assay. Spermatozoa taken from a cryopreserved pool of fertile donor specimens served as a control in the hemizona assay, and were used to assess the ability of the zona pellucida to bind spermatozoa. The mean ± SD number of spermatozoa attached to the hemizona was 107 ± 42. The binding capacity of different oocytes from the same cohort varied substantially (coefficient of variation = 28%). Age was not found to be correlated with the number of spermatozoa bound to the zona pellucida (r = -0.02; P > 0.1). It was concluded that female age has no role in the ability of the human zona pellucida to bind spermatozoa. Key words: female age/spermatozoa/zona binding.