Is early embryo cleavage a factor to increase success in all types of ICSI indications?.

PURPOSE: The authors aimed to compare early embryo cleavage with pregnancy rates in intracytoplasmic sperm injection/embryo transfer (ICSI/ET) cycles due to male infertility or tubal factor infertility (TFI). MATERIALS AND METHODS: 412 ICSI/embryo transfer cycles undergoing treatment for over two years were prospectively analyzed; 337 of the cycles were due to male infertility, whereas 75 suffered from tubal factors. Non-early cleaved (NEC) embryos were used for ET in 271 male factor and 67 tubal factor cycles, whereas early cleavage embryos were used for embryo transfer in 66 male factor and eight tubal factor cycles. RESULTS: In 66 out of 337 cycles (19.58 %) in male factor group and in eight out of 75 tubal factor cycles (10.66%), early cleavage (EC) embryos were obtained (p = 0.069). The clinical pregnancy rate was significantly elevated in EC subgroup (34.8%) compared to NEC subgroup (20.6%) (p = 0.015) in the male factor infertility group. The clinical pregnancy rate was non-significantly elevated in EC subgroup (37.5%) compared to NEC subgroup (23.8%) (p = 0.410) in the TFI group. CONCLUSIONS: The authors found that the implantation and pregnancy success of EC embryos vary with the therapeutic indication. The success rate would be low even with usage of EC embryos in untreated cycles of TFI.

Investigation of the association between the outcomes of sperm chromatin condensation and decondensation tests, and assisted reproduction techniques.

The main purpose of this prospective study is to examine possible influences of abnormalities of sperm nuclear condensation and chromatin decondensation with sodium dodecyl sulphate (SDS)-EDTA on outcomes of intrauterine insemination (IUI) or intracytoplasmic sperm injection (ICSI) cycles. Semen samples from 122 IUI and 236 ICSI cycles were evaluated. Before semen preparation for IUI or ICSI, basic semen analysis was performed and a small portion from each sample was spared for fixation. The condensation of sperm nuclear chromatin was evaluated with acidic aniline blue, followed by sperm chromatin decondensation by SDS-EDTA and evaluation under light microscope. Ongoing pregnancy rate was 24% and 26.2% in the IUI and ICSI groups respectively. The chromatin condensation rate was significantly higher in the ongoing pregnancy-positive group compared to the negative group, both in IUI (P = 0.042) and ICSI groups (P = 0.027), and it was positively correlated with ongoing pregnancy rate in both IUI and ICSI groups (P = 0.015, r = 0.214 and P = 0.014, r = 0.312 respectively). Chromatin decondensation rates were not significantly different in neither of the groups. These results indicate that IUI and ICSI outcome is influenced by the rate of spermatozoa with abnormal chromatin condensation. Sperm chromatin condensation with aniline blue is useful for selecting assisted reproduction techniques (ART) patients.

Comparison of follitropin alfa and urinary gonadotropins in IVF cycles.

AIM: The aim of this study was to compare human menopausal gonadotropins with recombinant follicle stimulating hormone-follitropin alpha following a long down-regulation protocol in intra cytoplasmic sperm injection cycles in our clinic, and to review the outcomes in the light of preceding studies. METHODS: This was a retrospective study. Among a total number of 2798 patients who had undergone IVF/ICSI applications, 579 eligible patients were included, and their data were evaluated retrospectively. Three hundred eighteen patients were treated with follitropin alpha and 255 patients were treated with hMG. Total units of follitropin alpha preparations used in ovulation induction, total number of meiois-2 phase oocytes, total number of used oocytes in ICSI cycle, fertilization rate and clinical pregnancy rates of both groups were analyzed. RESULTS: Mean duration of stimulation was longer in the group of patients treated with rFSH-α compared to the second group of patients treated with hMG (8.88 days and 8.55 days, respectively; P<0.05). The number of transferred embryos were 3.08 and 2.68 for patients treated with follitropin alpha and hMG, respectively (P<0.05). Clinical pregnancy rates were %28 and %33 in the groups of patients treated with follitropin alpha and hMG, respectively. Even though a greater clinical pregnancy rate was noted in the hMG group, there was no statistically significant difference between the two groups (P>0.05). CONCLUSION: Our results indicate that there is no statistically significant difference between follitropin alpha and human menopausal gonadotropin in terms of the clinical pregnancy rates.

Sperm selection based on motility in polyvinylpyrrolidone is associated with successful pregnancy and embryo development.

The aim of this study was to investigate whether spermatozoon motility in polyvinylpyrrolidone (PVP) is associated with better embryo development and pregnancy rates in ICSI cycles. A total of 123 primary ICSI treatment cycles were included in this study. Semen samples were tested for motility before ICSI procedure in PVP. Within 3 min, the presence or absence of motility was recorded. Sperm functions were examined by the aniline blue (AB) chromatin condensation test and the hypoosmotic swelling test, and the chromatin stability was evaluated by inducing its decondensation with sodium dodecyl sulphate and ethylenediaminetetraacetic acid (EDTA). Fertilisation and embryo scoring were evaluated. Fifty (64%) of 78 women conceived in the PVP (+) group; and 12 (26%) of 45 women conceived in the PVP (-) group; the pregnancy rate was significantly higher in the PVP (+) group (P = 0.003). Semen parameters were observed to be similar in both groups. The mean number of total embryos obtained in ICSI procedure and transferred grade 1 embryos were significantly higher in PVP (+) group (P = 0.01 and P = 0.003 respectively). The presence of sperm motility in PVP is associated with increased pregnancy rate, higher percentage of good quality embryos, sperm chromatin condensation and decondensation.

Assessment of human sperm morphology by strict criteria: comparison of wet preparation versus stained with the modified Diff-Quik method.

Routine semen examination remains an important tool for the diagnosis and treatment in human subfertility. Of all semen parameters, sperm morphology seems to be one of the most powerful indicators of a man's fertilizing potential in vitro and in vivo. Lack of standardization of sperm morphology assessments remains the main reason for the usefulness of this parameter. The aim of this study was to analyze the agreement between the wet-stained preparations versus those stained with modified Diff-Quik for sperm morphology. A total of 100 unselected semen samples from infertile couples were analyzed. Sperm morphology was evaluated with unstained specimens and following modified Diff-Quik staining according to the strict (Kruger classification) criteria by two different examiners (intralaboratory blind assessment). Mean percentages of morphologically normal spermatozoa were identical on wet and stained preparation slides (4.79 vs. 4.61, p >.05). Wide divergence of results was found with respect to the percentage of sperm with head and midpiece defects with the two different preparations (p >.001). The percentage of sperm tail defects was similar in both methods (p >.05). Simple linear regression analysis between the two methods revealed very good correlation for the morphologically normal spermatozoa (r =.83), but poor correlation for the sperm head, midpiece, and tail defects (r =.25,.25, and.28, respectively). Wet preparation is suitable only for the morphologically normal spermatozoa, but to determine the percentage of the defective spermatozoa, staining the smear is recommended.

Sperm-preparation techniques for men with normal and abnormal semen analysis. A comparison.

OBJECTIVE: To compare two commonly used sperm-preparation techniques, density gradient centrifugation and swim-up procedures, with respect to their effects on acrosome reaction (AR), hypoosmotic swelling (HOS) and nuclear maturity in men with abnormal and normal semen analyses. STUDY DESIGN: In accordance with World Health Organization criteria, 23 men with abnormal (group I) and 20 men with normal (group II) semen analyses were included in a prospective, controlled study. Each semen specimen was divided into aliquots in order to assess AR, HOS and nuclear maturity, determined with acridine orange staining, in both raw and processed semen samples using the density gradient centrifugation and swim-up techniques. RESULTS: Initial semen samples in group I revealed diminished AR, HOS and nuclear maturity rates in comparison to those in group II. In group I, density gradient centrifugation improved AR, HOS and nuclear maturity rates more than did swim-up. However, in group II it improved only the AR; HOS rates were better than with swim-up. There was a significant positive correlation between sperm concentration and HOS rate in raw semen samples from group I. In the same group, motility and morphology correlated with the nuclear maturity rate but not with AR and HOS rates. Semen samples with better motility (> 20%) or morphology (> 25%) showed better nuclear maturity rates (> 50%) in men with abnormal semen analyses. Motility had a sensitivity of 77% and specificity of 90% in predicting nuclear maturity. Morphology had similar sensitivity but lower specificity (70%). CONCLUSION: Density gradient centrifugation is superior to the swim-up technique in improving AR, HOS and nuclear maturity rates in men with abnormal semen analyses. However, when only nuclear maturity rate is taken into account, the swim-up technique seems to be sufficient for selecting spermatozoa in men with normal semen analyses. The nuclear maturity rate also correlates with sperm morphology and motility.

Evaluation of the fetal kidney maturation by assessment of amniotic fluid alpha-1 microglobulin levels.

OBJECTIVE: To evaluate the fetal renal maturation by assessment of amniotic fluid microproteins and to show these proteins originate from fetal urine. STUDY DESIGN: Amniotic fluid proteins (total protein, albumin, high molecular weight protein-HMWP, low molecular weight protein-LMWP, alpha(1)-microglobulin and beta(2)-microglobulin) were determined in 39 pregnant women at delivery and by amniocentesis in 30 pregnant women. These values were compared with first urine values of neonates with the same gestational age. RESULTS: Albumin was the largest protein component in the amniotic fluid. LMWP showed an increase in the amniotic fluid until the end of the second trimester; and as pregnancy advanced a progressive decrease occurred in parallel to fetal renal maturation. After 26 weeks' gestation, a strong correlation was identified between LMWP levels and alpha(1)-microglobulin, and between LMWP and beta(2)-microglobulin. No significant difference was detected between LMWP levels in the first urine of the neonates and in amniotic fluids. CONCLUSION: Microproteins in the fetal urine are of fetal origin. Fetal renal maturation can be evaluated by measuring microproteins in the amniotic fluid. Fetal renal maturation is best reflected by alpha(1)-microglobulin.

Does leukocytospermia associate with poor semen parameters and sperm functions in male infertility? The role of different seminal leukocyte concentrations.

To investigate the effect of leukocytospermia on standard semen analysis and sperm function tests such as acrosome reaction, hypoosmotic swelling, antisperm antibody binding and cervical mucus penetration, a prospective clinical study was performed. Two hundred and nineteen male infertility patients undergoing investigation and treatment were included in the study. There was a significant association between acrosome reaction positivity and leukocytospermia according to WHO (World Health Organization) criteria. Increased hypoosmotic swelling test score, higher sperm concentration and enhanced acrosome reaction were closely related to leukocytospermia. When the patients were divided into subgroups according to seminal leukocyte concentrations, acrosome reaction and hypoosmotic swelling were observed to be higher in semen samples with higher leukocyte concentrations compared to those with low seminal leukocyte concentration. In addition, higher sperm concentrations were observed in semen samples with increased leukocyte levels compared to semen samples with low leukocyte levels. These results suggest that leukocytospermia may have a favorable effect on some sperm functions at seminal leukocyte concentrations between 1 and 3x10(6)/ml.

Antibiotic therapy in men with leukocytospermia.

OBJECTIVE: In this prospective study, we aimed to determine the efficacy of doxycycline and doxycycline plus ceftriaxone for the treatment of asymptomatic men with leukocytospermia. METHOD: Seventy men were included in this randomized and placebo-controlled study. White blood cell (WBC) concentrations were determined by peroxidase assay during the routine semen analysis. Twenty-four of 70 men with leukocytospermia were randomized as control group and administered placebo (group I), 25 received doxycyline alone (group II), and 21, doxycycline plus ceftriaxone (group III). Doxycycline, 100 mg, was given twice a day for ten days and ceftriaxone, 1 g, in two doses for only one day. After the treatment, semen analyses were repeated. RESULTS: After the treatment there was a significant decrease in WBC counts in groups II and III when compared with group I (P < .05). Both antibiotic regimens were found to be equally effective. However, the time needed for resolution of leukocytospermia (approximately 4 weeks) was similar between the control and treatment groups. CONCLUSION: Although both antibiotic regimens significantly, and equally, improved the white blood cell counts in men with leukocytospermia, they failed to treat the leukocytospermia, i.e., to bring the count below the limit of one million WBC/mL. Therefore, it is doubtful that antibiotic therapy should be recommended for asymptomatic men with leukocytospermia.

Effects of indoleamines on sperm motility in vitro.

Indoleamines are products of the pineal gland and are postulated to play an antigonadotrophic role in the reproductive system of mammals. In humans, indoleamines have been localized in tissue fluids such as plasma, serum and cerebrospinal fluid. Because indoleamines exhibit antigonadotrophic properties, the authors examined whether these agents cause inhibitory effects on sperm motility. In this study, time and dose-dependent inhibition of sperm motility by indoleamines was observed. Furthermore, the presence of indoles in incubation medium decreased sperm velocity. These data suggest that the presence of high doses of indoles in reproductive fluids may inhibit sperm motility and velocity.

Effect of a pineal indolamine, 5-methoxyindole-3-acetic acid, on the estrous cycle and reproductive organs of female Wistar albino rats.

It has been suggested that the pineal gland has a specific role in the control of cyclic sexual activity in rats. One or more of the compounds isolated from this gland have been considered to be possible anti-fertility agents. In this study, the effect of different doses of 5-methoxyindole-3-acetic acid (5-MIAA) on the estrous cycle and the reproductive organs of female rats was studied. Daily injections of either 100 or 200 micrograms of 5-MIAA significantly prolonged the estrous cycles of mature female rats. After injections of 5-MIAA, an increase in diestrous smears was observed; in addition, the animals had estrous cycles without proestrous phase. Following the injections of 200 micrograms of 5-MIAA, a significant increase in uterine weight, plasma 17 beta-estradiol level and in the number of follicles were observed, and after injections of 5-MIAA (50, 100, 200 micrograms) a medium level of plasma progesterone was obtained. Thus it is concluded that 5-MIAA has an effect on the ovulation process of female rats.