Arabin cervical pessary in women at high risk of preterm birth: a magnetic resonance imaging observational follow-up study.

OBJECTIVE: To help elucidate the mechanism of action of the Arabin cervical pessary in pregnancies at high risk for preterm delivery. METHODS: Cervical length and uterocervical angle were evaluated in relation to gestational age in 198 pregnancies not at high risk for preterm birth that underwent clinical fetal magnetic resonance imaging (MRI). Additionally, in 73 singleton pregnancies at high risk for preterm birth, an Arabin cervical pessary was placed at 14-33 weeks' gestation. We performed MRI of the cervix immediately before and after placement and at monthly follow-up until removal of the pessary. In a subgroup of 54 pregnancies with a short cervix and pessary placement at 17-31 weeks' gestation, the uterocervical angle and cervical length at follow-up were compared with the preplacement values. RESULTS: In pregnancies not at high risk for preterm birth, the uterocervical angle did not vary, but cervical length showed a significant decrease with gestational age (r = -0.15, P < 0.05). Among the high-risk patients, the cervical pessary was successfully placed at first attempt in 60 (82.2%) and by the second attempt in 66 (90.4%), remaining well positioned until removal. In five patients we failed to place the pessary after two attempts, in one patient the pessary dislodged during follow-up and in one case the pessary was partly placed in the external cervical canal and triggered labor the next day. Among the subgroup of 54 patients, the median uterocervical angle immediately after pessary placement was significantly more acute than that prior to placement in the 46 (85.2%) who delivered after 34 weeks (132° vs 146°, P < 0.01), but was unchanged in the eight patients who delivered before 34 weeks (143° vs 152°, P > 0.05). CONCLUSION: In patients at high risk for preterm delivery, correct placement of the Arabin cervical pessary should be checked immediately; this can be performed quickly and easily using MRI. This study provides some evidence that, in singleton pregnancies with a short cervix, a cervical pessary delays birth through a mechanical effect on the uterocervical angle.

Canine hip dyslasia: the significance of the Norberg angle for healthy breeding.

OBJECTIVES: To assess the reproducibility of the Fédération Cynologique Internationale (FCI) classification over time, and between assessors and to evaluate the benefit of computer-assisted measurement of the Norberg angle and of its percentile ranking by breed for the diagnosis of canine hip dysplasia, and for the selection of couples for breeding. METHODS: During the time period of 2006 to 2010, 5094 hip radiographs were evaluated according to the FCI rules, and 4400 have been submitted for statistical analysis. A system of computer-assisted image analysis (Digimizer(®) , MedCalc Software Ltd, Mariakerke, Belgium) was used to measure the Norberg angle. The Norberg angle value of individual dogs was expressed as percentile rank by breed. RESULTS: The agreement between individual assessors was highly significant (P<0·001), but there were important variations over time of the ratio of classes A or B. The Norberg angle and the percentile rank accurately discriminated between dogs with or without canine hip dysplasia, with positive and negative likelihood ratios of 6·31 and 0·21, respectively, for the Norberg angle at criterion value of 102·2°, and 4·21 and 0·18 for the percentile rank at criterion value of the 25th percentile. CLINICAL SIGNIFICANCE: The percentile rank of the Norberg angle may be a valuable tool for breeding selection.

Combined conventional/antioxidant "Astaxanthin" treatment for male infertility: a double blind, randomized trial.

AIM: To evaluate the treatment of male infertility with a strong natural antioxidant, in addition to conventional treatment. METHODS: Using a double blind, randomized trial design, 30 men with infertility of > or =2 months and female partners with no demonstrable cause of infertility received conventional treatment according to the guidelines of the World Health Organization (WHO), and either a strong antioxidant Astaxanthin 16 mg/day (AstaCarox, AstaReal AB, Gustavsberg, Sweden) or placebo for 3 months. The effects of treatment on semen parameters, reactive oxygen species (ROS), zona-free hamster oocyte test, serum hormones including testosterone, luteinizing hormone (LH), follicle stimulating hormone (FSH) and Inhibin B, and spontaneous or intrauterine insemination (IUI)-induced pregnancies were evaluated. RESULTS: ROS and Inhibin B decreased significantly and sperm linear velocity increased in the Astaxanthin group (n = 11), but not in the placebo group (n = 19). The results of the zona-free hamster oocyte test tended to improve in the Astaxanthin group in contrast with the placebo group, though not reaching statistical significance. The total and per cycle pregnancy rates among the placebo cases (10.5 % and 3.6 %) were lower compared with 54.5 % and 23.1 % respectively in the Astaxanthin group (P = 0.028; P = 0.036). CONCLUSION: Although the present study suggests a positive effect of Astaxanthin on sperm parameters and fertility, the results need to be confirmed in a larger trial before recommending Astaxanthin for the complementary treatment of infertile men.

Hormonal regulation of inhibin B secretion by immature rat sertoli cells in vitro: possible use as a bioassay for estrogen detection.

The influences of follicle-stimulating hormone (FSH), gonadal steroids, and culture time were studied in relation to inhibin B production by Sertoli cells of immature rats cultured in vitro. Sertoli cell-enriched cultures were established from 18-day-old rats and were maintained in medium supplemented with insulin, transferrin, and epidermal growth factor at 34 degrees C. A recently developed ELISA for the measurement of inhibin B was used to assess the effects of recombinant human FSH (rh FSH), testosterone (T), and estradiol (E2) on inhibin B production and accumulation in the culture media of Sertoli cell-enriched cultures and to optimize the cell culture system to serve as a bioassay for the detection and quantification of estrogens and estrogenlike substances. Prolonging the incubation time (24, 48, or 72 hours) of Sertoli cells with control medium without rh FSH, T, or E2 resulted in a time-dependent increase of inhibin B production. Incubation with rh FSH (1, 2.5, 5, or 10 U/L) caused a dose- and time-dependent increase of inhibin B production by Sertoli cells (but not by cultured Leydig cells), reaching a plateau at 5 U/L rh FSH. Addition of T in concentrations of 2.88, 5, or 50 ng/ml to medium without rh FSH and E2 significantly lowered the daily production rate of inhibin B (P < 0.05). In contrast, addition of E2 (0.01 and 0.1 ng/ml) caused a dose-responsive increase in inhibin B production after 24 and 48 hours. The relative increment of inhibin B production induced by E2 was maximal after 24 hours in the presence of 2.5 U/L rh FSH (acting synergistically) and in the absence of T. When these conditions are implemented, the Sertoli cell culture system may serve as a bioassay for estrogenic substances, and it may reflect the possibly harmful effect they may have on spermatogenesis.

The Multi Unit Activity analyzer: a Windows based hardware-software system for low cost, high speed analog to digital data conversion, data acquisition and window discrimination.

The Multi Unit Activity analyzer is a hardware-software package for multi-purpose, two-channel data acquisition, with a computer dependent maximal digitizing frequency selectable from 1 to 27,000 s(-1) on both channels simultaneously. The hardware is connected to an IBM compatible PC through one of the serial ports (standard RS 232 interface). Software was developed to view digitized signals and record or read them on or from the harddisk. The program can also perform amplitude based window discrimination on the raw signal, on-line or during replay. The system is used for recording and analyzing multi unit activity from neuronal tissue in our electrophysiology lab but it can be applied in a variety of other settings. Basic programming routines are available that allow customized data acquisition.

White blood cells cause oxidative damage to the fatty acid composition of phospholipids of human spermatozoa.

The lipid composition of the sperm membrane has been shown to exert a significant effect upon the functional quality of spermatozoa. We have studied the effect of induced peroxidation and of the presence of polymorphonuclear white blood cells (WBCs) on the fatty acid composition of the phospholipids of human spermatozoa. The spermatozoa were fractionated by a discontinuous Percoll gradient in two fractions (47% and 90% Percoll). Induced peroxidation of spermatozoa was assessed by determining the production of thiobarbituric acid reactive substances (TBARS), mostly malondialdehyde, after incubation with ferrous sulphate and sodium ascorbate as a promoter of peroxidation. TBARS production after induction of peroxidation was correlated with the abundance of polyunsaturated fatty acids (PUFA)(r = 0.68, p < 0.0001), with the double bond index (r = 0.72, p < 0.0001), and with the oxidative potential index (r = 0.73, p < 0.0001) of fatty acids of phospholipids. In comparison with samples containing > 1 x 10(6) WBCs/mL, those with < 1 x 10(6) WBCs/mL contained higher proportions of PUFA (90% Percoll, p < 0.05; 47% Percoll, p < 0.05), total omega 3 fatty acids (90% Percoll, p < 0.05; 47% Percoll, p < 0.001), docosahexaenoic acid (90% Percoll p < 0.05; 47% Percoll, p < 0.05), and double bond index (90% Percoll, p < 0.05; 47% Percoll, p < 0.001). In addition, mean melting point was significantly lower (90% Percoll, p < 0.05; 47% Percoll, p < 0.001) in samples with < 1 x 10(6) WBCs, indicating higher membrane fluidity. The increase of TBARS production by spermatozoa after incubation with the xanthine-xanthine oxidase system and/or ferrous sulphate as promoter of peroxidation was associated with a significant decrease of PUFA. Incubation of spermatozoa with WBCs, with or without activation by phorbol ester, decreased the PUFA (p < 0.05). Also, TBARS production was increased (p < 0.01) after activation of WBCs with phorbol ester. Our data provide evidence that oxidative stress induced by WBCs has a damaging effect on the polyunsaturated fatty acids of sperm phospholipids which may result, amongst other effects, in decreased membrane fluidity.

The fatty acid composition of phospholipids of spermatozoa from infertile patients.

The lipid composition of the sperm membrane has a significant effect upon the functional characteristics of spermatozoa. In the present study we investigated the fatty acid (FA) composition of subpopulations of spermatozoa separated on a discontinuous Percoll gradient (47:90%) and the FA composition of phospholipids (PL) of sperm heads and tails in both normal and abnormal semen samples. In normozoospermic samples, polyunsaturated fatty acids (PUFA) represented 34.0 +/- 1.3 (mean +/- SE, mole %) and 25.6 +/- 1.2% of total FA of PL of the 47 and 90% Percoll fractions respectively. Docosahexaenoic acid (22:6omega3, DHA) contributed to more than 60% of total PUFA. DHA was significantly lower in both the 47% (P < 0.05) and the 90% (P < 0.01) Percoll fractions of oligozoospermic samples and in the 90% Percoll layer of asthenozoospermic samples (P < 0.01), compared with normozoospermic samples. The omega6/omega3 ratio was significantly increased in both Percoll fractions of samples with oligozoospermia (47%, P < 0.001 and 90%, P < 0.001) or with asthenozoospermia (47%, P < 0.05 and 90%, P < 0.001) compared with normozoospermic samples. The oxidative potential index (OPI) of spermatozoa recovered from the 47% Percoll layer was significantly higher (P < 0.0001) than of those recovered from the 90% Percoll. Mean melting point (MMP), an index of membrane fluidity, was significantly lower in head than in tails (P < 0.01) of spermatozoa, and also in both the 47% (P < 0.01) and 90% (P < 0.001) Percoll fractions of normozoospermic samples in comparison with oligozoospermic samples. The MMP was significantly higher (P < 0.05) in samples of patients with idiopathic oligo/asthenozoospermia, varicocele, and male accessory gland infection (MAGI). These differences in FA composition of PL in subpopulations of human spermatozoa, and in their heads and tails may be related to sperm maturity and to differences in physiological function.

The relation between reactive oxygen species and cytokines in andrological patients with or without male accessory gland infection.

The presence of various cytokines, namely hepatocyte growth factor (HGF), interleukin-1 receptor antagonist (IL-1 RA), and interleukins (IL-1 alpha, IL-6, and IL-8), as well as the production of reactive oxygen species (ROS) was investigated in seminal plasma of fertile and infertile patients in order to evaluate the possible value of measuring these substances for the diagnosis of male accessory gland infection, and to assess the possible relationship between oxidative stress and cytokines during leucocytospermia and male accessory gland infection (MAGI). Our findings indicate that all of the measured cytokines seem to be produced locally as well as by white blood cells (WBC) and that, due to the presence of higher numbers of WBC, accessory gland infection may exert a deleterious effect on sperm quality through the production of ROS and/or of particular cytokines such as IL-1 alpha, IL-1 RA, and IL-8. The most specific marker for a sensitivity of 95% in discriminating between cases with or without MAGI is the measurement of IL-6 in seminal plasma. In the absence of WBC several cytokines are constitutively produced and correlate with sperm concentration (HGF, IL-8), alpha-glucosidase (IL-6), and gamma-glutamyltransferase activity (HGF). The measurement of these cytokines in semen may provide clinically useful information for the diagnosis of male accessory gland infection, as well as in the absence of WBC where it can provide information about certain mechanisms of male reproductive function and dysfunction.

The possible meaning of transferrin and its soluble receptors in seminal plasma as markers of the seminiferous epithelium.

Transferrin (Tf) and soluble transferrin receptors (S-Tf-R) were measured by enzyme immunoassay in seminal plasma of 130 semen samples. The mean concentration of S-Tf-R in cases with normozoospermia was 10.4 IU/ml (95% confidence interval: 9.5-11.3) and it was significantly lower in patients with oligozoospermia (6.6, 95% CI: 5.8-7.5, P < 0.001), asthenozoospermia (8.5, 95% CI: 5.5-10.7, P < 0.05), azoospermia of primary testicular origin (7.9, 95% CI: 6.1-9.6, P<0.05) and post-vasectomy samples (5.9, 95% CI: 5.4-6.9, P < 0.001). The concentration of S-Tf-R in post-vasectomy samples was lower than that in patients with azoospermia of primary testicular origin (P < 0.05; positive likelihood ration = 7 at value of 8.3 IU/ ml). S-Tf-R was positively correlated with motile sperm concentration (r = 0.50, P < 0.0001), percentage motility (r = 0.38, P < 0.001), percentage of normal forms (r = 0.43, P < 0.001), sperm linear velocity (r = 0.42, P < 0.001), and ATP concentration (r = 0.67, P < 0.0001). Follicle stimulating hormone (FSH) was found to be negatively correlated with the concentrations of both Tf (r = -0.31, P < 0.05) and of S-Tf-R (r = -0.45, P < 0.01). The mean concentration of Tf in seminal plasma was 50.4 micrograms/ml (35.9-67.2) in samples with normozoospermia (n = 22), and the concentration was significantly lower in patients with oligozoospermia (P < 0.05), azoospermia of testicular origin (P < 0.001), and post-vasectomy samples (P < 0.001). Seminal Tf was correlated with motile sperm concentration (r = 0.36, P < 0.001), percentage of motile spermatozoa (r = 0.25, P < 0.05), linear velocity (r = 0.24, P < 0.05) and ATP concentration (r = p.44, P < 0.001). The concentration of Tf was positively correlated with that of S-Tf-R both in cases with spermatozoa present (r = 0.66, P < 0.001), and in cases with azoospermia of testicular origin (r = 0.51, P < 0.05) but not in vasectomy cases. It is concluded that S-Tf-R in seminal plasma is a marker of spermatogenesis and may give information on the presence or absence of spermatogenetic cells in cases with azoospermia. Further investigations are needed to assess its usefulness for clinical practice.

Deterioration of sperm quality in young healthy Belgian men.

We have retrospectively analysed the sperm characteristics of 416 consecutive healthy young men who presented themselves in the past 19 years as candidate sperm donors. Ejaculate volume increased slightly (P = 0.067), and average sperm concentration decreased (P = 0.035) by 12.4 x 10(6)/ml over the observation period, so that sperm count per ejaculate remained unchanged (P = 0.91). In contrast, sperm morphology (r = - 0.23, P < 0.0001), rapid progressive motility (r = - 0.42, P < 0.0001) and total motility (r = - 0.33, P < 0.0001) presented an important and time-related decrease. When a quadratic model was used rather than a linear one to analyse the data on rapid progressive motility, there appeared to have been no further decline since 1990. The average proportion of spermatozoa with normal morphology decreased from 39.2% in the period 1977-1980 to 26.6% in 1990-1995 (P < 0.0001), and the mean percentage of spermatozoa with rapid progressive motility decreased from 52.7 to 31.7% (P < 0.0001). The percentage of candidate donors with sperm characteristics below the 5th percentile cut-off value of a normal fertile population increased from 13 to 54% during the observation period (P < 0.0001). Since the technique of semen analysis has remained essentially unchanged in-so-far as has been practically possible, as has the method of recruitment of candidate sperm donors, the observed deterioration of sperm characteristics is considered to reflect degeneration of sperm production among men aged between 20 and 40 years.

Statement on the general reduction in sperm quality.

Over a period of 17 years, 360 young healthy men presented themselves as candidate donors for the sperm bank of the University Hospital of Ghent. Semen characteristics of these men were assessed using unaltered methods, and analysis was performed on the first sample delivered before any donor selection was performed. Sperm motility and morphology, but not the total sperm count per ejaculate, exhibited a highly significant decrease. Over 40% of candidate donors since 1990 exhibited subnormal sperm characteristics, as against only 5% of the group investigated before 1980.

Varicocele: indications for treatment.

Varicocele is detected in about one-third of men with subfertile semen quality. Treatment by means of transcatheter embolization using the tissue adhesive Histoacryl improves Sertoli cell function with increased concentrations of transferrin in semen, and a significant increase in sperm concentration, normal morphology and linear velocity. Failure to attain pregnancy after varicocele treatment occurs if pre-treatment sperm quality is normal, or in the case of azoospermia with elevated blood FSH levels or obstruction, or when there is severe coincidental pathology impairing fertility of the male or female partner. In all other cases, the poor results of varicocele treatment reported in some studies can only be explained by inadequate techniques. Varicocele detection and embolization treatment at puberty or during adolescence should be advocated to prevent deterioration of testicular function.

MedCalc: a new computer program for medical statistics.

In recent years, the use and abuse of statistics in the medical literature has extensively been reviewed. Amongst others, the importance of the P-value has been challenged and the use of misleading graphics, including 3-dimensional displays, has been criticized. The ease of access to more complex statistical procedures, since the introduction of several statistical software packages for personal computers, has been identified as one of the factors involved in the misuse of statistics. Therefore, we have developed a new computer program that includes those statistical procedures commonly encountered in the medical literature and in statistical textbooks for medical researchers. More complex statistical analyses are not implemented in the software. If researchers with limited statistical training require more sophisticated statistical analyses, they should refer to a statistician, not to a more complete statistical software package.

Does treatment with testosterone undecanoate improve the in-vitro fertilizing capacity of spermatozoa in patients with idiopathic testicular failure? (results of a double blind study).

Seventy-seven couples in whom conventional in-vitro fertilization (IVF) had remained unsuccessful because of low fertilization rate and abnormal sperm characteristics were given either testosterone undecanoate 120 mg/day, or placebo during 3 months, after which a new IVF treatment was applied under identical technical conditions. There were no significant changes in sperm characteristics among the treated and placebo couples and the fertilization rate showed a similar increase after treatment in both groups. No significant difference in pregnancies occurred, with 32% pregnancies in the placebo controls and 17% among couples treated with testosterone undecanoate. It is concluded that testosterone undecanoate intake does not improve sperm characteristics, or the in-vitro fertilizing potential, or pregnancy rate over those observed in the placebo controls in cases with primary idiopathic testicular failure.

The effective cumulative pregnancy rate of different modes of treatment of male infertility.

The clinical efficacy of conventional and advanced methods of treatment was assessed in 814 couples with infertility due to a male factor. The monthly and effective cumulative rate of ongoing or term pregnancies was calculated during 4712 couple-months. Treatment of varicocele by transcatheter embolization, resulting in 3.9% pregnancies per cycle and an effective cumulative pregnancy rate of 41% after 1 year, is more effective than counselling and timed intercourse (9% pregnancies after 12 months). Intrauterine insemination (IUI) of washed spermatozoa produced 17% pregnancies in the initial 4 months, but the success rate of the subsequent cycles (1.7% per cycle) was not different from that of the controls. In vitro fertilization (IVF) resulted in 16% pregnancies per attempt, but the effective cumulative pregnancy rate was only 31% in 12 months due to the long interval between treatment attempts and the high drop-out rate. With subzonal microinjection of sperm, the fertilization rate was higher (71%) than with regular IVF (29%) but both the pregnancy rate per attempt (9%) and the effective cumulative pregnancy rate (17% after 12 months) were low. The 10th percentile of sperm characteristics (cut-off values) of successful cases showed intrauterine insemination to be advantageous in cases with a lower percentage of spermatozoa with progressive motility (9%) than in the controls (15%). The cut-off value of sperm morphology in IVF (4%) is lower than that of IUI (8%) and of the controls (9%), but higher than that of subzonal insemination (1%). Treatment strategy must be defined selecting or combining conventional and assisted reproductive technology for each individual couple with male factor infertility.

Comparison of different methods for the investigation of antisperm antibodies on spermatozoa, in seminal plasma and in serum.

Since there is no gold standard for the diagnosis of immunological infertility, comparison between different methods and of their results with biological tests for the detection of antisperm antibodies must be used in defining the most reliable and clinically relevant method. We have evaluated the results of direct (n = 100) and indirect tests in serum (n = 140) and seminal plasma (n = 100) using the SpermMAR and Immunobead (IB) techniques for the detection of sperm antibodies of the immunoglobulin (Ig) classes G, A and M, and we have compared the results with those of the tray agglutination test and the adenosine triphosphate release cytotoxicity test. Results indicate the higher specificity and sensitivity of the SpermMAR results for IgG and IgA, as compared to the outcome of the IB test. It appears that the IB test for IgA may detect non-specific antibodies in serum. Little information is obtained from the tray agglutination test in seminal plasma, and the indirect SpermMar test for IgG in seminal plasma was the only independent variable associated with the outcome of the former test. Since the SpermMAR tests for IgG and IgA are more accurate and biologically relevant, as well as easier to perform than the respective IB tests, the former must be considered the method of first choice.