Combination of hypoosmotic swelling/eosin Y test for sperm membrane integrity evaluation: correlations with other sperm parameters to predict ICSI cycles.

The objective of our study was to evaluate the accuracy of the combination of hypoosmotic swelling (HOS) and eosin Y (Ey) exclusion tests to predict the ICSI cycles' outcome and its correlations with other sperm parameters. The functional and structural integrity of sperm membrane was evaluated with the combined HOS/Ey test in 95 ICSI cycles and the results were correlated with other sperm parameters, including concentration, motility, strict morphology, and total motile sperm count. The combined HOS/Ey test was evaluated for the prediction of the ICSI cycles' outcome parameters including fertilization, cleavage, and pregnancy rates. The HOS/Ey test presented significant relationships with concentration, motility, and strict morphology (p < 0,0001) but it couldn't predict the fertilization, cleavage, and pregnancy outcomes of ICSI cycles. The combined HOS/Ey test has strong correlations with motility and strict morphology parameters of sperm samples but is not sufficiently sensitive to estimate the outcome of ICSI cycles.

Ultrastructural changes on sperm after extracorporeal shock wave lithotripsy in patients with distal ureteral stone.

Theoretically, ESWL can cause several side effects on the male reproductive system. We determined here the long-term effects of ESWL on sperm with transmission electron microscopy (TEM) in patients with distal ureteral stone. Fifteen men with stones in the distal ureter applied to our clinic formed the group of study. The other 15 men with renal or upper ureter stones formed the group of control. The ESWL sessions, including maximum 19 kW energy level and 3000 shock waves, were performed with Siemens Lithostar (electromagnetic; Siemens Medical Systems, Erlangen, Germany) lithotriptor. We examined the semen samples from all patients on the day before and 90 days after ESWL. The semen samples were examined with transmission electron microscopy (TEM) to detect ultrastructural changes on the day before and 90 days after ESWL. All the statistical analyses were realized with SPSS 10.0 (SPSS Inc., Chicago, USA) statistical package program. When the control and study groups were compared for initial and day 90 sperm concentration and motility, a significant decrease was found in the study group. Although there was no important anomaly in the control group, we determined some damage on sperm structure in 5 patients (33.3%) who are in the study group 3 months after ESWL. It can reduce sperm concentration and motility permanently. It can also cause severe ultrastructural defects on sperm after a long term period in patients with lower ureteral stone. Therefore, we suggest other treatment modalities for young men with distal ureteral stones to prevent the development of male infertility.

Effects of phototherapy on newborn rat testicles.

Phototherapy is the most widespread treatment for lowering bilirubin concentration in neonates. In the routine, phototherapy has some side effects including skin eruption, fluid loss, abdominal distention, mild hemolysis and mild thrombocytopenia. The aim of the study was to investigate the possible mutagenic and gametocidal side effects of 72 h continuous phototherapy on the rat testicle. We observed decreases in spermatogonia numbers per tubule (S/T values), tubular fertilization index (TFI) and sperm sertoli cell index (SSCI), which are the most reliable methods in estimating future fertility potential, due to sensitivity to phototherapy. The differences between study and control groups for S/T, TFI and SSCI values were statistically significant (p = 0.008, p = 0.02 and p = 0.004, respectively). There were significant differences in seminiferous tubule diameters between the control and study groups (p < 0.005), but no significant difference in DNA index values between the control (0.66 +/- 0.12) and study (0.59 +/- 0.05) groups (p > 0.05). As a conclusion, phototherapy seems to have some side effects on the newborn rat testicle. Further studies with larger groups, designed for investigation of the effects of phototherapy on seminiferous tubules, may give more beneficial results.

Effects of exposure to new inhalational anesthetics on spermatogenesis and sperm morphology in rabbits.

The effects of chronic exposure to new inhalation anesthetics (sevoflurane and isoflurane) on spermatogenesis and sperm morphology were examined in 23 rabbits, randomly divided in 3 groups. Rabbits received 20 exposure hours (four hours/day x 5 days), as follows: group I: 2.3% (1.2 MAC) sevoflurane + 2L/min oxygen, group II: 1.3% (1.2 MAC) isoflurane + 2L/min oxygen, and group III (control): 2L/min oxygen. Semen was collected on the 12th, 19th, 26th, 33rd, and 41st days of exposure. Sperm concentration, motility and morphological changes were evaluated. On the 41st day, testicular biopsies were taken and observed with light microscopy. Sperm concentration and motility significantly decreased in the sevoflurane and the isoflurane groups, compared to control. There were no significant changes in the control group. It is concluded that chronic exposure to the new inhalational anesthetics had negative effects on spermatogenesis and sperm morphology.

Low sperm recovery from the undescended testis with testicular sperm extraction in postpubertal cryptorchids: preliminary report.

As postpubertal cryptorchid testes are generally atrophic and have high malignancy risk, orchiectomy is recommended as the treatment method. However, a few case reports advocated orchiopexy due to finding sperm afterwards. The present clinical study aimed at determining the possibility of finding spermatozoa in postpubertal cryptorchid testis with testicular sperm extraction (TESE). Initially testicular biopsy and later TESE was performed in the undescended testis of 22 unilateral postpubertal cryptorchids and in one of the testes of three bilateral postpubertal cryptorchids. Histopathological assessment of testicular biopsy specimen showed that three patients had maturation arrest and 22 had seminiferous tubular atrophy. By TESE, spermatozoa were found in only one specimen of 25 testes. Our results show that the expectancy to find spermatozoa in postpubertal cryptorchid testis is extremely low by current sperm recovery methods and orchiectomy is still the accurate method of treatment.

Pronuclear synchronization and nuclear morphology of mature and in vitro matured oocytes in the rat: an ultrastructural study.

The objective of this study was to evaluate synchronous and asynchronous pronucleus (PN) formation and the related patterns of juxtapositional nucleolus (n) formation in immature (prophase I [PI] and metaphase I [MI]) and mature (metaphase II [MII]) oocytes after fertilization, both ultrastructurally and at the level of light microscope. A single dose of 15 IU gonadotrophin was injected subcutaneously to twenty four 26-wk-old, female Wistar rats to induce ovulation. Human chorionic gonadotrophin (4 IU) was administered 40 h later, and after 4-6 h the ovaries were dissected, and the oocytes were aspirated. A total of 214 rat oocytes were classified according to a maturation index as follows: group I, 80 PI oocytes; group II, 50 MI oocytes; and group III, 84 MII oocytes. Immature oocytes were in vitro matured for 18-36 h. Spermatozoa were acquired by microepididymal sperm aspiration and processed using swim-up technique. Intracytoplasmic sperm injection was performed on mature oocytes after 2 h of incubation and on in vitro matured (IVM) oocytes 4 h after maturation. Pronuclear synchronization [both pronucleases (PNs) centrally located, equal sized, with equal numbers and sizes of juxtapositional nucleoli (Nn)] was observed in fertilized oocytes. Asynchronous PN formation (diversity between male and female PNs, related to dimensions, localization, and the number of Nn) in groups I, II, and III was found in 75, 86, and 47% of preembryos, respectively. There was a significant difference of synchronous pronuclear formation between mature and IVM oocytes (P < 0.05). In IVM oocytes, asynchronous PN formation is high, and juxtapositional pronucleolar patterns are observed to be low by transmission electron microscope (TEM).

The mast cells in semen: their effects on sperm motility.

This study was conducted to evaluate any possible association between mast cells and sperm concentration, morphology, and motility. The study comprised 400 patients who had applied for semen analysis. To evaluate mast cells, 6 smear slides were prepared for each subject and stained with 1% toluidin blue-pyronine (pH 4). The slides revealing any mast cells were labeled as mast+. Concentration and motility was evaluated through a Makler chamber. Kruger's strict criteria were used in morphometric analysis. The mean age of 86 mast+ cases (21% of total patients) was 31+/-6.7; progressive sperm motility rate was 33+/-21.2. The mean concentration was 32+/-30.2 x 10(6)/mL, and normal sperm percentage was 11.8+/-6.5. Progressive sperm motility rate in the mast- cases were 53+/-25. The mean age of mast cell+ patients was higher than that of mast cell- patients (t=3.57, p<.001), while they had lower sperm concentration (p>.05) and lower normal morphologic sperm rate (t=2.26, p<.024), compared to mast cell- patients. The relation between mast cell+ and mast cell- cases and sperm progressive motility was statistically significant (t=6.44, p<.001). It was concluded that sperm parameters were negatively affected by mast cells.

Oocyte membrane maturation and oocyte-sperm relationship: transmission electron microscopy study.

The success of in vitro maturation (IVM) depends greatly on the acquisition of immature oocytes. Immature oocytes in prophase I (PI) and metaphase I (MI), aspirated after controlled ovarian hyperstimulation, were incapable of fertilization, leading to a lower fertilization rate. Therefore, they must be evaluated on a fine structure level for their in vitro maturation (IVM) processes and their relationship with sperm. Oocyte membrane maturation and oocyte-sperm relationship were studied using transmission electron microscopy. A total of 55 human oocytes obtained from 20 patients at various times and 83 oocytes obtained from the dissected ovarians of female Wistar rats were used for transmission electron microscopy (TEM) evaluation. Despite being in either prophase I and metaphase I or in metaphase II, the oocytes were not fertilized after 48 h of incubation. At the various stages of maturation between PI and MII, the number and the size of microvilluses on the oocyte membrane increased as MII approached and decreased after full maturation. Oocyte activation was related to oocyte membrane maturation and has an effect on the oocyte sperm penetration.