Rotational digital subtraction carotid angiography: technique and comparison with static digital subtraction angiography

To compare the efficiency of sperm preparation between the two-layer Percoll gradient and mini-Percoll methods, 50 normal and 33 abnormal semen samples from male partners of infertile couples were studied. The number of recovered spermatozoa, percentage of motility, percentage of normal morphology, and their survival at 24 and 48 hours were assessed. Both Percoll gradient techniques resulted in a significantly higher percentage of motility and percentage of normal morphology compared with the original semen samples (p < 0.0001). The two-layer Percoll gradient showed a higher sperm recovery than the mini-Percoll method (p < 0.001), but the latter resulted in a higher percentage of motility (p > 0.001) and a higher sperm survival rate at 24 hours (p < 0.05) than the former, regarding normal semen samples. These differences did not appear with abnormal semen samples when analyzed as a group. Considering each of the abnormal parameters separately, sperm recovery was significantly higher after the two-layer Percoll gradient in the case of astheno- and teratozoospermia (p < 0.05), but sperm survival at 48 hours was higher after the mini-Percoll gradient in the case of teratozoospermia (p < 0.05). It is concluded that both the two-layer Percoll gradient and mini-Percoll method can be used effectively for sperm preparation. The former yields a higher sperm recovery, but the latter should be considered regarding teratozoospermic samples and semen samples of very low volume.     

Spermatozoa selection by the swim-up procedure and two-layer percoll gradient centrifugation

The efficiency of the swim-up procedure and two-layer Percoll gradient centrifugation in procession of spermatozoa was assessed in ejaculates from 47 infertile men. A significantly higher total number of spermatozoa was harvested from Percoll gradients than from the swim-up procedure, the loss rates in concentration being -13.6 +/- 6.4% and -70.8 +/- 5.8%, respectively (p < 0.0001). Recovery in per cent motility was significantly higher after the Percoll gradient than after the swim-up procedure (34.8 +/- 10.2% versus -10.4 +/- 17.2%, p < 0.05). No significant difference was noted between the mean motility grades of the final solutions obtained by the two methods (2.7 +/- 0.2 and 2.0 +/- 0.4, respectively, p > 0.05). When evaluation was conducted within three initial fresh sample concentration categories such as severe oligospermia (lower than 5 x 10(6)/ml), moderate oligospermia (5 to 10 x 10(6)/ml) and mild oligospermia (higher than 10 x 10(6)/ml), the Percoll technique recovered significantly higher number of spermatozoa than the swim-up procedure through all concentration categories (p < 0.05 for each range). Despite being statistically insignificant, Percoll gradients produced final spermatozoa pools with higher per cent motility and motility quality within all concentration ranges. The results suggest that the Percoll gradient centrifugation should be the preferred selection method regardless of the initial fresh sample concentration.     

Improved motile sperm recovery by a hyperosmotic Percoll gradient

PURPOSE: Our purpose was to investigate whether a new, relatively hyperosmotic Percoll gradient, Enhance-S, can improve total motile sperm recovery rates compared with the commonly used Percoll gradient Perception. METHODS: Semen specimens from each of 17 donors were divided into two equal aliquots. One part was washed using Percoll Perception, while the other was prepared using Percoll Enhance-S. RESULTS: Compared to the unwashed specimen, sperm motion characteristics (motility and velocity) improved significantly after Percoll separation using either the Perception or the Enhance-S gradient. There was no difference in motility or velocity in spermatozoa recovered after wash with either of the two preparations. However, the total motile sperm recovery was significantly higher using the Percoll Enhance-S gradient than with the Percoll Perception gradient (P < 0.0024). CONCLUSION: The new Percoll Enhance-S gradient provides significantly more total motile sperm than the Percoll Perception gradient.     

Effect of follicular or oviductal fluids on movement characteristics of bovine sperm during capacitation in vitro

Mammalian sperm exhibit characteristic motility changes associated with capacitation. Movement characteristics of bovine sperm incubated in noncapacitating (control, medium alone), capacitating (oviduct fluid, nonluteal, and luteal), or capacitating, acrosome reaction inducing (follicular fluid) conditions were investigated using a computer-assisted automated semen analysis system. Sperm were incubated up to 4 hours in a modified Tyrode's medium (control), 20 and 60% nonluteal (NL) or luteal (L) oviduct fluid (ODF), or 20 and 60% follicular fluid (FF). Relative to sperm incubated in control medium, motility of sperm treated with ODF or FF had increased linearity and vigorous motility. Sperm incubated in 60% ODF or FF showed a small decrease in mean trajectory/path straightness and velocity over time compared to 20% fluid treatments and control. Frequency distribution graphs were symmetric for 20% NL- and L-ODF treated sperm. However, 20% FF and 60% ODF and FF treatments had distributions skewed to the left, indicating smaller values for lateral head displacement (ALH) and curvilinear velocity (VCL). Median values for ALH and VCL were determined for control-treated sperm, and subtracted from individual sperm values for all treatments to estimate deviation from control, designated ALHc and VCLc. Three-dimensional plots of ALHc, VCLc and corresponding frequency indicated shifts in peak patterns for fluid-treated sperm compared to control sperm. Incubation in 20% ODF and FF resulted in peak shift for ALH and VCL values; yet, little change in peak position was observed in sperm incubated in 60% ODF and FF.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of various methods of processing human cryopreserved-thawed semen samples

We compared the efficacy of various methods of processing cryopreserved-thawed samples for the recovery of functionally adequate spermatozoa as assessed by the response to the sperm stress test (SST), an index of temperature activated sperm membrane lipid peroxidation, and immediate and delayed changes in sperm viability and motion parameters. Donor semen samples (n = 28) were cryopreserved-thawed and divided into six equal parts, one part was used as control and the remaining parts were used to compare five methods of sperm processing as follows: direct Percoll gradient processing, washing by one-step or stepwise addition of the washing medium followed by Percoll processing, and washing by one-step or stepwise addition of the washing medium. Additional samples (n = 10) were evaluated for the immediate and delayed (6 h at 37 degrees C) impact of one-step and stepwise washing (without Percoll separation). Compared with wash-only methods, samples processed using Percoll had a significantly higher SST score (P = 0.001), motility, rapid spermatozoa (>50 microm/s), curvilinear velocity and motility index (P < 0.001). Comparing various Percoll methods, direct Percoll processing resulted in the highest number of motile spermatozoa recovered (P < 0.00001) and a higher SST score based on curvilinear velocity (P = 0.001). Stepwise washing gave a significantly higher number of motile spermatozoa (P < 0.001) but with a significantly lower SST score based on the concentration of motile spermatozoa (P = 0.001), motility (P = 0.001) and motility index (P = 0.01). Sperm viability and motion parameters after 6 h of incubation showed no difference between one-step and stepwise washing. In conclusion, compared with wash-only methods, Percoll processed samples resulted in the recovery of spermatozoa with superior quality as assessed by SST and motion analysis. One-step washing of the samples gave an overall comparable recovery compared to the samples prepared stepwise. Having significantly higher SST scores, similar viability and the maintenance of motility, one-step washing may be a better method of processing thawed samples than the stepwise washing.     

Rationale for cigarette smoking and for mentholation preference in cocaine- and nicotine-dependent outpatients

The effects of sauna exposure on sperm movement characteristics and other semen parameters were evaluated using computer-assisted sperm analysis (CASA). A significant (p < 0.01) decrease in average path velocity (VAP), curvilinear velocity (VCL) and amplitude of lateral head displacement (ALH) was found after exposure to sauna for 2 weeks. The altered parameters returned to their original values within 1 week after cessation of sauna exposure. Mean values for semen volume, sperm count, percentage motility, sperm morphology and sperm penetration assay (SPA) were not statistically different during and after sauna, when compared to the corresponding control values. The results suggest that increasing scrotal temperature by sauna causes a reversible decrease in sperm movement parameters.     

Variation in recovery of motile sperm after preparation by a simple Percoll gradient technique

PURPOSE: Numerous techniques have been used to prepare sperm for assisted reproduction technology. Density-gradient centrifugation with Percoll is becoming a method of choice. This study reviewed the results of a simple two-layer discontinuous Percoll gradient. METHODS: We reviewed retrospectively results obtained from 208 semen specimens prepared for in vitro fertilization by a discontinuous Percoll gradient. RESULTS: Overall results (mean +/- SD) were a recovery of 21 +/- 16% of total sperm, recovery of 38 +/- 30% of motile sperm, and a motility in the final sperm preparation of 88 +/- 10%. Specimens with higher initial concentrations had higher motility in the final preparation. Higher total recovery and higher motility in the final preparation were found for specimens with a higher initial motility. CONCLUSIONS: This simple two-layer Percoll technique is rapid and inexpensive and reliably produces a final sperm preparation with desirable characteristics. Even though specimens with poor initial parameters yielded final preparations with excellent characteristics, fertilization rates were still related to the initial semen parameters.     

Synthesis and antibacterial evaluation of novel water-soluble organic peroxides

Various compounds have been used in the attempt to improve sperm motility, including pentoxifylline (PF), a methylxanthine derivative. It has been postulated that PF, being a phosphodiesterase inhibitor, increases sperm kinematic parameters and the number of spermatozoa exhibiting hyperactivated motility by raising the intracellular content of cAMP, a molecule involved in the generation of sperm energy. However, it has not been clarified whether the biological effects of PF on sperm motility correlate with its ability to increase intracellular cAMP levels. To examine this relationship, the kinematic parameters, hyperactivation, and intracellular cAMP content were evaluated in motile spermatozoa, obtained by discontinuous Percoll gradient and swim-up from 21 normozoospermic semen samples, incubated without and with PF for 0, 1, 2, and 4 h. PF increased beat cross frequency after 1 and 2 h of incubation, curvilinear velocity and lateral head displacement (ALH) after 4 h, and hyperactivation after 1, 2, and 4 h, and decreased linearity (LIN) after 1 h of incubation. The intracellular cAMP content of spermatozoa incubated with PF increased at all time-points examined. Both intracellular cAMP content and increase in hyperactivation in response to PF decreased with the length of incubation. In the absence of PF, cAMP content was unchanged and was correlated significantly only with ALH and the percentage of spermatozoa with hyperactivated motility. Following incubation with PF, cAMP content correlated with hyperactivation and all sperm kinematic parameters, with the exception of LIN and straightness. These findings suggest that the beneficial effects of PF on sperm kinematic parameters and hyperactivation are related to its ability to increase intracellular cAMP content.     

An auto-controlled study in in-vitro fertilization reveals the benefit of Percoll centrifugation to swim-up in the preparation of poor-quality semen

The efficiency of spermatozoa prepared by swim-up or by Percoll centrifugation was assessed in an in-vitro fertilization programme on 71 semen samples of a well-defined quality [total number of type A (WHO criteria) motile spermatozoa]: category I (n = 21) with > 100 x 10(6), II (n = 31) with 15-100 x 10(6), III (n = 11) with 5-15 x 10(6) and IV (n = 8) with < 5 x 10(6) type A motile spermatozoa. Oocytes were inseminated 4 h after oocyte retrieval, alternately with spermatozoa derived from swim-up and Percoll preparation. Both selection procedures resulted in a significantly higher (P < 0.001) percentage motility as compared to fresh semen. For low-quality samples (III and IV), however, swim-up was more effective in selecting highly motile (P = 0.004) and morphologically normal spermatozoa (P < 0.05). For high-quality samples, this difference might have been masked by introducing a swim-up step to remove Percoll particles. Regardless of the initial sperm quality, the mean fertilization rate was significantly higher (P = 0.003) when Percoll-treated spermatozoa were used for insemination (51.3 versus 37.8%). For semen of groups I and II, no difference in fertilization capacity was observed according to the sperm preparation method. Despite the lower percentage motility and normal morphology for the Percoll compared to the swim-up treatment in groups III and IV, fertilizing capacity was significantly (P < 0.001) in favour of this selection method (65.3 versus 26.5% in group III, 47.6 versus 11.6% in group IV).(ABSTRACT TRUNCATED AT 250 WORDS)     

Microsatellite deletion mapping on chromosome 10q and mutation analysis of MMAC1, FAS, and MXI1 in human glioblastoma multiforme

The aim of this study was to determine the relationship between calcium ionophore A23187-induced acrosome reaction (AR) and sperm fertilizing ability. Semen samples remaining after preparation for standard IVF were studied in 109 patients who had sperm concentrations > or =20 x 10(6)/ml. Ionophore-induced AR was performed on motile spermatozoa selected by centrifugation on a Percoll gradient. Semen analysis was performed using standard methods. Patients with higher (>50%, n = 76) fertilization rates had significantly higher ionophore-induced AR than patients with lower (<50%, n = 33) fertilization rates (49 +/- 14 versus 38 +/- 21%, P < 0.05). When the data from all patients were analysed by logistic regression, only the percentage sperm motility in insemination medium and ionophore-induced AR were significantly related to fertilization rates. Similar results were also obtained when the data from a subgroup of patients with poor (<15% normal) sperm morphology were analysed. However, when patients with normal sperm morphology > or =15% were analysed separately, only sperm count and the percentage of spermatozoa with progressive motility in semen were significantly related to fertilization rates. In conclusion, ionophore-induced AR was significantly related to fertilization rates in vitro mainly in patients with teratozoospermic semen. Tests for ionophore-induced AR may provide additional information about sperm fertilizing ability but may not indicate specific defects of the physiological AR.     

Enhancement of motility by treating spermatozoa with an antioxidant solution (Sperm-Fit) following ejaculation

Oxygen radical generation is known to be detrimental to sperm function, especially motility, through the lipid peroxidation of the membranes. Generation of reactive oxygen species can be induced by leukocyte contamination, sperm centrifugation and the presence of abnormal spermatozoa with excess residual cytoplasm. This study aims to evaluate the effect on sperm motility of incubation in an antioxidant-containing solution, during liquefaction and centrifugation. Thirty semen samples were each divided into two equal parts: one mixed with Tyrode's solution, the other with a salt solution containing antioxidants (Sperm-Fit; Ellios Bio-Media, Paris, France). All the procedures were identical in the two groups. The ratio of leukocytes to spermatozoa was significantly correlated with the motility after liquefaction and after a 24 h incubation in routine in-vitro fertilization (IVF) medium and with the number of motile spermatozoa recovered after Percoll preparation. Moreover, when this ratio was > or = 0.2, all motility parameters were lowered. Incubation with Sperm-Fit allowed a higher percentage of motility after Percoll preparation when the ratio was > or = 0.2 (48 +/- 5% versus 41 +/- 6% for Sperm-Fit and Tyrode's solution respectively; P < 0.05) and a greater number of motile spermatozoa recovered after Percoll preparation, whatever the ratio (3.2 +/- 1.0 x 10(6) versus 2.4 +/- 0.7 x 10(6) for Sperm-Fit and Tyrode's solution respectively when ratio > or = 0.2; 18.1 +/- 3.4 x 10(6) versus 14.4 +/- 2.9 x 10(6) for Sperm-Fit and Tyrode's solution respectively when ratio < 0.2; P < 0.05). These results show that incubation with antioxidants during liquefaction and centrifugation increases recovery of motile spermatozoa.     

Post-thaw sperm motility, cAMP concentration and membrane lipid peroxidation after stimulation with pentoxifylline and platelet-activating factor

Earlier studies have demonstrated that pentoxifylline (PTX) and platelet-activating factor (PAF) can significantly improve the motion parameters of post-thaw human spermatozoa. This study has investigated the effects of PAF, PTX and their combination on cyclic adenosine monophosphate (cAMP) concentrations and membrane lipid peroxidation (LPO) in post-thaw human spermatozoa. Washed spermatozoa from normal volunteers (n = 10) were cryopreserved in Test-yolk buffer using a standard protocol. After 2 weeks the sperm samples were thawed, washed and incubated with either 1 microM PAF, 3 mM PTX or 0.5 microM PAF plus 1.5 mM PTX. Video sequences were recorded at 0, 30, 60, and 120 min for analysis of sperm motion parameters using the Cell Track Sperm Analysis System. Concentrations of cAMP were assessed by radioimmunoassay, and LPO levels were measured by malondialdehyde-thiobarbituric acid reactivity. Our studies indicate a time-course stimulatory effect with overall maximal stimulation observed in samples treated with the combination of PAF and PTX. The maximal stimulation of percentage motility compared to control was observed at 60 min in samples treated with PAF, PTX, or PAF plus PTX. PAF plus PTX stimulated straight-line velocity (VSL), curvilinear velocity (VCL) and lateral head displacement (ALH) after 30 min incubation. The primary effect of PAF was observed on VSL, while the main effect of PTX was on VCL. cAMP concentrations were 3-fold higher than controls in samples treated with PTX or PAF plus PTX. cAMP concentrations in PAF-treated samples did not differ significantly from controls. No significant differences were observed between any groups for LPO.(ABSTRACT TRUNCATED AT 250 WORDS)     

Discontinuous Percoll gradient preparation for donor insemination: determinants for success

The use of cryopreserved donor spermatozoa for insemination has become necessary to decrease the risk from sexually transmitted infectious diseases. Lower fecundity rates have been reported with this practice. Efforts have been applied to increase success, including identification of those sperm characteristics which correlate with increased fecundity. Data from in-vitro fertilization have revealed sperm morphology, motility and zona pellucida binding as important sperm parameters. Discontinuous Percoll gradient preparation yields a high concentration of motile spermatozoa. Using this preparation for thawed donor spermatozoa, we have identified post-preparation motility and progression as factors associated with increased fecundity. Consideration should be given to screening sperm donors with a freeze-thaw Percoll gradient preparation prior to acceptance into a donor bank.     

Comparison of sperm separation methods: effect on recovery, motility, motion parameters, and hyperactivation

OBJECTIVE: To compare the Enhance (Percoll; Conception Technologies, San Diego, CA) and PureSperm (Gen X International, Madison, CT) sperm preparation methods with respect to recovery (percentage of motile sperm), motility (%), path and progressive velocities (microm/s), and hyperactivation (%). DESIGN: Comparison of sperm processing methods. SETTING: University medical center-based clinical andrology laboratory and infertility program. PATIENT(S): Twenty-five men who presented for semen analysis. INTERVENTION(S): Each of 25 semen specimens were divided and each aliquot was prepared using two different density gradient centrifugation methods. MAIN OUTCOME MEASURE(S): The motile sperm recovery, percent motility, motion parameters, and percent hyperactivation were measured for each semen specimen (n=25) before and after separation with the use of the two methods. RESULT(S): There was no difference in the percent motility and motile count between specimens prepared with Enhance (Percoll) and PureSperm and fresh specimens. Statistically significant differences were found (fresh versus test) in the velocities and in hyperactivation (PureSperm only), and no differences were found between the processing methods. CONCLUSION(S): PureSperm appears to be as effective as Percoll (Enhance) for the recovery of good, progressively motile sperm for use in IUI or other assisted reproductive techniques.     

Inhibin B in men with normal and disturbed spermatogenesis

Inhibin, a dimeric gonadal glycoprotein, inhibits the production and/or secretion of follicle stimulating hormone (FSH). The major species currently recognized are inhibin A (alphabeta A subunit) and inhibin B (alphabeta B subunit). In men, inhibin B seems to be the physiologically important form of inhibin. Therefore we measured serum inhibin B using a new two-site immunoenzymatic assay in 14 men (mean +/- SEM age, 34.5 +/- 0.7 years) with sperm counts >20 x 10(6)/ ml, in 35 men (mean +/- SEM age, 36.4 +/- 1.3 years) with oligozoospermia (sperm count <20 x 10(6)/ml) and in men with azoospermia (three orchidectomized men, three men with Klinefelter's syndrome, 10 men with Kallmann's syndrome). We compared inhibin B concentrations with serum FSH and sperm concentrations. In men with normal sperm concentrations (44.7 +/- 6.4 x 10(6)/ml), the concentration of inhibin was 223 +/- 18 pg/ml and of FSH 5.0 +/- 0.7 IU/l; in patients with low sperm concentrations (3.7 +/- 0.8 x 10(6)/ml), the concentration of inhibin B was 107 +/- 12 pg/ml and of FSH 12.2 +/- 1.5 IU/l. In all patients, except those with hypogonadotrophic hypogonadism, the relationship between inhibin B and FSH concentrations was inverse (r = -0.69, P < 0.0001). In all patients the sperm concentration was positively correlated with inhibin B concentrations (r = 0.70, P < 0.0001) and negatively correlated with FSH concentrations (r = -0.37, P < 0.01). We conclude that inhibin B may be a marker of exocrine testicular function and could offer improved diagnosis and treatment modalities for male infertility.     

The paradoxical effects of pentoxifylline on the binding of spermatozoa to the human zona pellucida

In the presence of pentoxifylline, human spermatozoa are induced to increase certain motion characteristics; however, the role of this drug in fertilization remains equivocal. In this study, the influence of pentoxifylline on one aspect of fertilization, that is sperm-zona binding, has been examined. Results from a fluorescence label competitive zona binding (CZB) test showed that spermatozoa exposed to a pentoxifylline challenge of between 0.1 and 5 mM, which was curtailed after 1 h by washing, had a decreased (P < 0.01) ability to bind to intact zona compared with control spermatozoa. The washing procedures also removed (decrease P < 0.01 compared with peak values) some of the enhanced motion characteristics induced by pentoxifylline. These results were in contrast with those obtained using experimental conditions that maintained an increased curvilinear velocity (VCL) and lateral head displacement (ALH) (increase P < 0.001 above baseline controls) in the continued presence of pentoxifylline. Using a hemizona binding (HZB) assay, 3 mM pentoxifylline increased (P < 0.001) sperm-zona binding almost 20% above zona binding with unexposed control spermatozoa. It was concluded that, in the presence of pentoxifylline, there is increased sperm binding to the zona pellucida; however, if the drug is removed by washing, the sperm binding to the zona is decreased in concert with the removal of the enhanced motion characteristics. The application of zona solubilization by acidic conditions in a microchamber enabled the precise determination of sperm numbers in both of the sperm-zona binding assays, and the results demonstrated that a wide variation in sperm numbers was observed in each test, with 63-580 spermatozoa bound in the CZB assay and 56-1340 spermatozoa bound on a hemizona.     

Usefulness of the acrobead test in evaluating human acrosome function in fresh and cryopreserved sperm

PURPOSE: Assays for the acrosome reaction are usually cumbersome and lack reproducibility. Accurate determination of acrosomal status is important in patients diagnosed with male infertility before proceeding with intrauterine insemination or in vitro fertilization. We determined the optimum capacitation time and acrosomal status of fresh semen specimens in normal fertile men with the Acrobead test, and whether the assay could be used to evaluate cryopreserved semen specimens. MATERIALS AND METHODS: Semen samples from 13 normal donors were divided, with half of the fresh ejaculate used for the Acrobead test and half cryopreserved for a minimum of 24 hours in liquid nitrogen before testing. Fresh and frozen specimens were prepared with the swim-up technique. Sperm concentration was adjusted to 4, 2, 1 and 0.5 x 10(6)/ml. in 4 wells of a 96-well tissue culture plate. Ten microl. polyacrylamide beads (1.5 x 10(6)/ml.) coated with anti-CD46 monoclonal antibodies (MH61 beads) were added to each well. The attachment of beads with acrosome reacted spermatozoa was scored after 0, 1, 3, 6 and 24 hours of incubation. Results were graded on a scale of 0 (no bead binding to the sperm) to 4 (complete attachment to the beads). Specimens with scores of at least 2 were considered normal. RESULTS: A score of at least 2 was noted in 3 of 13 fresh specimens (15.3%) at 1, 9 (69.2%) at 3, 11 (84.6%) at 6 and 13 (100%) after 24 hours. However, a significantly greater number of frozen specimens (8 of 13, or 62%) had a score of 2 or more at 1 hour of incubation and 100% bead attachment to sperm occurred at 3 hours. CONCLUSIONS: Our results indicate that in fresh semen specimens an incubation period of 6 to 24 hours can be used to screen individuals who present with normal sperm characteristics but have slow acrosome reactions. Early acrosome reaction observed in cryopreserved specimens indicates that these spermatozoa may have membrane damage and leakage of acrosome contents as a result of the freeze-thaw process. The Acrobead assay is a simple and objective test that can be used at a clinical andrology laboratory to evaluate the acrosomal status of fresh but not frozen human spermatozoa.     

Sperm morphology using strict criteria after Percoll density separation: influence on cleavage and pregnancy rates after in-vitro fertilization

The main purpose of the study was to evaluate the use of sperm morphology assessment by strict criteria on the post-Percoll separated spermatozoa used for oocyte insemination in an in-vitro fertilization programme. This study included a consecutive unselected series of 213 oocyte aspirations in 159 women. In 177 aspirations the patient had tubal infertility and in 36 unexplained infertility. Data have been analysed from 197 aspirations where the semen sample used for insemination had a normal sperm concentration (> or = 20 x 10(6)/ml). A total of 1413 oocytes were aspirated, resulting in 863 oocytes which were fertilized and cleaved (cleavage rate 61%). In all, 492 pre-embryos were transferred in 193 cycles, resulting in a pregnancy rate of 42% per transfer. Sperm morphology evaluation using strict criteria showed that Percoll separation significantly increased the percentage of sperm cells with normal morphology from 7.7 to 11.3%. Sperm morphology analysis showed that Percoll separation decreased the number of sperm samples in the 'poor prognosis pattern' group from 31 to 13% and increased the number of sperm samples classified as 'normal' from 16 to 33%. After Percoll separation the poor prognosis pattern group had a cleavage rate of 46%, which was significantly lower than in the good prognosis pattern and the normal groups. However, the poor prognosis pattern group had a significantly higher pregnancy rate than the normal group (P < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)     

Treatment of human spermatozoa with follicular fluid can influence lipid content and motility during in vitro capacitation

In order to evaluate the role of human follicular fluid (HFF) on the fertilizing capacity of spermatozoa, we studied the effect of HFF on the lipid composition and on the movement characteristics of human spermatozoa. Spermatozoa (spz) from normospermic patients were prepared with a discontinuous Percoll gradient and incubated in Ménézo B2 medium with or without a supplement of 20% HFF (HFF-Percoll spz and B2-Percoll spz respectively) for 2 and 24 h. After 2 h HFF incubation, percentage progressive motility, straight line velocity (VSL), and amplitude of lateral head displacement (ALH) were improved in HFF-Percoll spz as compared to B2-Percoll spz (P < or = 0.05). After a longer incubation period (24 h), lipid changes appeared in HFF-Percoll spz with lower levels of cholesterol (P = 0.02) and phospholipids (P = 0.05). No modification of the cholesterol/phospholipid ratio after 2 and 24 h of incubation in either B2-Percoll spz or HFF-Percoll spz was observed. Such decreases in lipid content of HFF-Percoll spz may be factors which could be taken into account as constituting part of membrane modifications during the capacitation process.     

Randomized comparison of intravenous immunoglobulin and methylprednisolone pulse therapy in children with newly diagnosed idiopathic thrombocytic purpura. The Danish ITP Study Group

Forty three children with newly diagnosed idiopathic thrombocytopenic purpura (ITP), platelet count (pl.c.) below 20 x 10(9)/l, and either clinically significant bleeding or failure to show a spontaneous platelet rise within three days of admission were randomly allocated to treatment with intravenous infusions of either immunoglobulin (IVIG) 1 g/kg or methylprednisolone (MPPT) 30 mg/kg on two consecutive days. Prompt induction of partial remission with pl.c. > 50 x 10(9)/l after 72 hours was seen in 21/23 given IVIG versus 10/20 given MPPT (exact p = 0.003); mean pl.c.s after 72 hours were 188 versus 77 x 10(9)/l (2p < 0.001). Poor responders were then given the alternative infusions in addition. After six days, complete remission with pl.c. > 150 x 10(9)/l was achieved in 16/23 versus 10/20 (p = 0.16). During six months follow-up, there were no significant differences regarding relapse rates or chronic course. Eleven children with relapse were crossed over to the alternative treatment arm: the estimated treatment effect in pl.c. after 72 hours was 134 x 10(9)/l in favour of IVIG. These results indicate that IVIG infusions may be preferable to high-dose corticosteroids as initial treatment for children with ITP.