Genitourinary cancer patients have worse baseline semen parameters than healthy sperm bankers.

BACKGROUND: While the spermatotoxic properties of cancer treatments such as chemotherapy and radiation therapy are widely recognized, the effect of malignancy itself on male fertility is not clearly understood. OBJECTIVES: To determine whether malignancy is associated with diminished semen quality prior to spermatotoxic treatment among sperm bankers. MATERIALS AND METHODS: Retrospective database review of de-identified records was obtained for all episodes of sperm banking performed at a cryobank from January 2004 to May 2017 for one of the following reasons: 'future use' (e.g., military deployment and gender reassignment); infertility; benign disease; and malignancy, further categorized as testicular, other genitourinary (GU), solid non-GU, hematologic, or unspecified. Dependent variables of interest were ejaculatory volume, sperm concentration, % motility, and total motile sperm count (TMSC), as well as post-thaw TMSC. RESULTS: A total of 1558 patients met the inclusion criteria. Multivariable regression analysis on log-transformed data controlling for age demonstrated decreased ejaculatory volume and sperm concentration, % motility, and TMSC in the infertility group as compared to the 'future use' group (p < 0.001). Testicular cancer was associated with decreased sperm concentration, TMSC, and post-thaw TMSC (p < 0.001); other GU malignancy was associated with decreased ejaculatory volume (p < 0.001). Benign disease, solid non-GU malignancy, hematologic malignancy, and unspecified malignancy were not associated with decreased parameters. DISCUSSION: In addition to sperm bankers with known fertility issues, sperm bankers with testicular and other GU malignancy had worse baseline semen parameters as compared to individuals pursuing banking for future use. These findings can inform patient counseling and consent prior to sperm banking and disease treatment. CONCLUSION: Individuals with testicular and other GU malignancy who banked spermatozoa before undergoing spermatotoxic therapy demonstrated worse baseline semen parameters as compared to individuals banking spermatozoa for non-medical reasons.

Sperm cryopreservation of transgender individuals: trends and findings in the past decade.

OBJECTIVES: Awareness and acceptance of transgenderism have increased in the last two decades. There is limited literature regarding the incidence and semen characteristics of transwomen banking spermatozoa. We sought to assess the incidence of sperm cryopreservation of transgender individuals compared with the cisgender population in the last 10 years. Semen parameters were also compared between the two groups. MATERIALS AND METHODS: We performed a retrospective analysis of sperm cryopreservation performed at a single center from 2006 through 2016. Using available data on indications for banking and prior hormonal therapy status, we isolated healthy transgender and cisgender cohorts for semen parameter comparison. Linear regression was used to compare the incidence trends. Semen parameters were compared using the generalized estimating equations method. The rates of semen parameter abnormality of each group were compared using chi-square test. Semen parameter abnormalities were defined using WHO 2010 reference values. RESULTS: We analyzed 194 transgender samples and 2327 cisgender samples for a total of 84 unique transgender sperm bankers and 1398 unique cisgender sperm bankers. The number of transgender sperm bankers increased relative to cisgender sperm bankers from 2006 to 2016. Following exclusion of cisgender sperm bankers with health issues that might impact semen quality and transgender sperm bankers with known prior hormonal therapy, we compared the semen parameters of 141 healthy cisgender sperm bankers and 78 healthy transgender sperm bankers. The transgender sperm bankers demonstrated lower sperm concentration, total motile sperm count, and post-thaw sperm parameters. The transgender sperm bankers also demonstrated a higher incidence of oligozoospermia. CONCLUSIONS: This is the largest report to date on the incidence of transgender sperm cryopreservation and comparison of semen characteristics with cisgender sperm bankers. The data reveal an increased incidence of transgender sperm banking as well as poorer semen parameters of transgender individuals compared with cisgender controls.

Decline in sperm count and motility in young adult men from 2003 to 2013: observations from a U.S. sperm bank.

Controversy exists regarding stability of semen quality over time with papers reporting decrease, increase or stable parameters in heterogeneous populations. The current study examined semen parameters of young adult men from 2003 to 2013 at an urban U.S. sperm bank. Semen parameters were analyzed before and after cryopreservation for a total of 9425 specimens from 489 individuals. Demographic information was obtained from a social and medical history questionnaire. Following 2-3 days abstinence, the specimens were collected at the laboratory and assessed by uniform technicians and techniques. The data were analyzed using generalized linear regression after adjustment for age, days of abstinence, for repeated samples, as well as by the Cochran-Armitage trend test. The within variability was accounted for by the repeated measures model. All p values were two-sided with p < 0.05 considered significant. There was a significant decline in sperm concentration (-3.55, 95% CI -4.87, -2.23; p < 0.001), total motility (-1.23, 95% CI -1.65, -0.82; p < 0.001), total count (-10.75, 95% CI -15.95, -5.54; p < 0.001) and total motile count (-9.43, 95% CI -13.14, -5.73; p < 0.001). There was no significant change in semen volume (0.03, 95% CI -0.02, 0.09; p = 0.2). The post-thaw total motility significantly (-2.30, 95% CI -2.72, -1.87; p < 0.001) decreased with time. Importantly, demographic and lifestyle factors were stable or improved over the study period. There was a decline in age (p(trend) = 0.003) and alcohol use (p(trend) = 0.005) and an increase in college GPA (Grade Point Average) (p(trend) = 0.02). BMI (p(trend) = 0.73), educational attainment (p(trend) = 0.2), race/ethnicity (p(trend) = 0.53), and lifestyle habits (weekly exercise, p(trend) = 0.21; smoking, p(trend) = 0.99; marital status, p(trend) = 0.85) remained constant. Uniform technicians and techniques over the study period make measurement bias unlikely. This report demonstrates a decline in semen quality among young adult men in the Boston area who were attending or completed a college education during the past 10 years, and requires further study.

Metal ions and human sperm mannose receptors.

Zinc and lead concentrations were measured in seminal plasma from fertile donors, infertile men with varicocoele and men undergoing work-ups for in vitro fertilization. Ejaculated spermatozoa from these subjects were incubated in vitro with various metal ions and/or dibromoethane and dibromochloropropane. Mannose receptor expression was correlated with metal and toxicant levels. Sperm distributions of potassium channels were compared with lead ions and calcium channels with zinc ions. Mannose receptor expression by capacitated spermatozoa increased linearly with seminal plasma zinc levels, and correlated inversely with lead levels. Cobalt had no effect on mannose receptor expression, but nickel had a concentration-dependent biphasic effect. Mannose receptor expression was not affected by dibromoethane and dibromochloropropane if the cholesterol content of the sperm membrane was high, but mannose receptor expression was decreased in low cholesterol spermatozoa by exposures below estimated permissive exposure limits. Potassium channels and lead ions co-localized over the entire head of human spermatozoa, while both calcium channels and zinc ions were confined to the equatorial segment of the head. Mannose receptor expression on the external surface of the human sperm plasma membrane is a biomarker for the effects of transition and heavy metals and organic toxicants on sperm fertility potential.

Seasonal variations and age-related changes in human sperm count, motility, motion parameters, morphology, and white blood cell concentration.

OBJECTIVE: To determine the presence of any seasonal variations and age-related changes in sperm parameters in andrology patients and fertile donors. DESIGN: Retrospective analysis. SETTING: University medical center andrology laboratory. PATIENT(S): The database of 2,065 semen analyses was retrospectively reviewed for the period of March 1, 1996, to October 31, 1998. INTERVENTION(S): None. MAIN OUTCOME MEASURES(S): The sperm count, motility, motile count, progressive straightline velocity, and percentage of rapid sperm were determined with the Hamilton-Thorne IVOS analyzer with standard setup parameters. RESULT(S): There were no significant seasonal differences in the patient's volume, sperm count, motility, motile count, whereas the percentage of rapid sperm and progressive straightline velocity were significantly lower in the spring. Correlation analysis of patient semen parameters versus age implied that as age increases there is a tendency for these semen parameters to decrease, whereas percent tail defects showed a significant positive correlation with age. CONCLUSION(S): Age-adjusted analyses of seasonal variations in andrology patient semen parameters showed significant seasonal variation in the percentage rapid motile sperm and straightline velocity, as well as the percent tail defects, percent immature sperm, and the percent tapered sperm. Such seasonal variations might prove to be clinically relevant and important when designing experimental protocols.

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.

Dose-response effects of gramicidin-D, EDTA, and nonoxynol-9 on sperm motion parameters and acrosome status.

Previous reports showed that gramicidin-D (G-D), a polypeptide with antiviral and antimicrobial properties, nonoxynol-9 (N9), a common spermicidal detergent, and EDTA, a Ca-Mg chelating agent, inhibited sperm motility and cervical mucus penetration. The purpose of this study was to determine the dose-response effects of G-D, N9, EDTA and G-D + EDTA on sperm motion parameters and acrosome status. Semen specimens from known fertile donors were subjected to computer-assisted semen analysis of motility, path velocity, progressive velocity, and hyperactivation prior to and after incubation with varying concentrations of gramicidin-D, EDTA and nonoxynol-9. Each specimen was also prepared for acrosome status using rhodamine isothiocyanate conjugated pisum sativum agglutinin (RITC-PSA). There was a significant decrease in motility by G-D, EDTA, G-D + EDTA, and N9 at all doses as compared to the fresh specimen. N9 completely immobilized all sperm at each dose. Progressive velocity and path velocity also decreased in a dose-response manner. Sperm hyperactive motility also significantly decreased in all groups. The majority of sperm remained acrosome intact following exposure to all doses tested, whereas N9 resulted in complete breakdown/release of the acrosomal contents. This study confirms previous reports that G-D, EDTA, and N9 significantly impair sperm motility and motion parameters. The effective 100% inhibitory concentration was seen only with N9, whereas G-D, EDTA, and G-D + EDTA resulted in incomplete impairment of sperm motion parameters. At the concentrations used, N9 demonstrated potent spermostatic activity. Gramicidin-D and EDTA should be further studied for their potential contraceptive spermostatic activity.

PIP: Gramicidin-D (G-D), a polypeptide with antiviral and antimicrobial properties, the spermicidal detergent nonoxynol-9 (N-9), and the Ca-Mg chelating agent EDTA have been shown in previous studies to inhibit sperm motility and cervical mucus penetration. This study utilized computer-assisted methods to investigate the dose-response effects of incubation with G-D, N-9, EDTA, and G-D plus EDTA on sperm motion parameters and acrosome status. Semen specimens were acquired within 30 minutes of ejaculation from six fertile US sperm donors. Compared to the fresh (untreated) specimen, there was a significant decrease in sperm motility produced by G-D, EDTA, G-D plus EDTA, and N-9 at all doses. Progressive and path velocity and sperm hyperactive motility also decreased in a dose-response manner in all groups. However, sperm immobilization was complete at the concentrations used only with N-9. The majority of sperm remained acrosome-intact after exposure to all tested doses of G-D and EDTA, but N-9 resulted in complete breakdown and release of the acrosomal contents. A combination of N-9 and G-D or N-9 and EDTA at lower doses might produce the desired inhibition of sperm motility without toxicity and this possibility should be investigated. At the present time, however, G-D or EDTA, alone or in combination, cannot be considered effective contraceptive agents.

Time course of spontaneous in vitro sperm acrosome reaction.

The acrosome reaction (AR) is an exocytotic process essential for sperm penetration of the zona pellucida and binding to the oocyte (DeJonge, 1994). Evaluation of in vitro AR can suggest fertility potential. The purpose of this study was to determine AR as a function of time after removal of sperm from the seminal fluid using a novel test, the Acrobeads test (Fertility Technologies, Natick, Massachusetts), which uses paramagnetic beads coated with MH61, a monoclonal antibody that binds to acrosome-reacted sperm. Specimens were acquired from known fertile donors (n = 9) and in vitro fertilization (IVF) patients (n = 8) with no apparent male factor on the day of the IVF and processed by a minipercoll wash at 30 minutes after ejaculation. An aliquot of washed sperm was then divided into two portions. The first was placed with the Acrobeads (according to the manufacturer's instructions) and assessed for bead binding after 6 and 24 hours with the beads. The second aliquot of washed sperm was held at room temperature for 24 hours, then exposed to the beads, with bead binding assessed at 6 and 24 hours later (30 and 48 hours after washing). The Acrobeads score was determined by assessing the binding of MH61 beads in each of four replicates with a resulting score of 1 (lowest) to 4 (highest). The mean (+/-SD) motility was 62.0% (7.5) at 6 hours, 52.3% (6.4) at 24 hours, 55.9% (10.4) at 30 hours, and 54.7% (8.4) at 48 hours after removal from the seminal fluid. At 6 hours after washing and exposure to the beads, the score was 0.077 (0.27) with a range of 0-1; one donor specimen gave a score of 1, while all others had a score of 0. At 24 hours postremoval from the semen, donor and patient sperm were positive for the test, with a mean score of 3.6 (0.65). The mean fertilization rate for the IVF patients was 64.4% (range 33-90). When sperm were held for 24 hours prior to the test, there was little or no bead binding 6 hours later (score of 0.46 +/- 0.77) and at 24 hours later (48 hours after washing) (mean score of 0.25 +/- 0.45). These data suggest that completion of the acrosome reaction occurs by 24 hours after removal of the sperm from the seminal fluid. Since the MH61 beads bind to specific residues on the inner acrosomal membrane, these data also suggest that following the acrosome reaction at 24 hours with removal of the outer acrosomal contents and acrosomal matrix, the inner acrosomal membrane may be modified in some way that does not allow MH61 beads to bind to the sperm.

Successful treatment of severe oligozoospermia with sperm washing and intrauterine insemination.

During the period January 1, 1991 through December 31, 1995, 258 patients, in whom motile sperm counts for insemination (postwash, processed) were 10.0 million motile sperm or less were seen in the andrology unit for sperm washing and intrauterine insemination (IUI). No significant female factors were noted on history; all female partners had patent Fallopian tubes and were ovulatory spontaneously or were treated by the referring gynecologist with clomiphene citrate, human menopausal gonadotropin (hMG), or follicle-stimulating hormone (FSH) ovulation induction in both anovulatory or ovulatory women. Of the total of 258 patients, 15 achieved a pregnancy in 284 cycles of IUI in which the inseminating motile-count was < 1.0 million motile sperm, resulting in a monthly fecundity (f) of 5.3%. The mean (+/-SD) motile count for IUI in this group was 0.61 (+/-0.29) million sperm, with a range of 0.19-0.95 million motile sperm. The initial motile count was 2.97 (3.2) million sperm, with a range of 0.2-12.81 million sperm. With inseminating motile counts of 1.0-10.0 million motile sperm, there were 83 pregnancies after 467 cycles of IUI, resulting in a monthly f of 17.8%. The mean (+/-SD) motile count for IUI in this group was 4.9 (+/-2.7) million motile sperm with a range of 1.0-9.9 million motile sperm. The initial sperm count in this group was 10.9 million (+/-7.1), with a range of 1.1-23.7 million motile sperm. These data suggest that acceptable pregnancy rates can be achieved with IUI, even in severely oligozoospermic specimens. Intrauterine insemination is less invasive and less costly than other assisted reproductive techniques. These data are supportive of IUI prior to attempting other more invasive and potentially costly reproductive technologies.

Comparison of the immunobead binding test (IBT) and immunospheres (IS) assay for detecting serum antisperm antibodies.

PROBLEM: The IBT is considered the gold standard of sperm antibody assays. This test uses polyacrylamide beads labeled with antiglobulins (anti-IgG, anti-IgA, and anti-IgM), which bind to the corresponding antibody on the sperm surface. The IS uses color-coded latex beads of uniform 3.0 microns size coated with the antiglobulins which can be viewed with brightfield light microscopy. The purpose of the present study was to compare the IBT and IS in an indirect test using human serum. METHOD: Serum specimens (n = 42) were tested for the presence of antibody isotypes IgG, IgA, and IgM to sperm using the standard protocol for IBT and IS. Donor sperm was washed in BWW with 5% BSA and diluted to a final concentration of 50 x 10(6) motile sperm/ ml. The sperm were incubated with a 1:10 dilution of test serum for 30 min to 1 h at 37 degrees C and then washed by three cycles of centrifugation. The sperm and beads (IBT, IS) were mixed on a glass slide, covered with a coverslip, and observed within 5 min. At least 100 motile sperm were counted and scored for bead binding. A specimen was considered positive if 20% or more of the sperm were coated with one or more beads. The data were analyzed using calculation of the non-parametric kappa statistic with correction for chance expected agreement, and by calculating the proportion of specific agreement between the two methods. RESULTS: The results are summarized in the following table: [table see text]; The IS was able to detect 94% of IgG antibodies, 91% of IgA antibodies, and 100% of IgM antibodies. One serum specimen was IgG negative by IS (14% binding), but positive by IBT (20%). A second serum specimen was IgA negative by IS (16%) yet positive by IBT (29%). There were no false positives with the IS assay. Of the IgM positives (five of six) occurred alone and not with IgG or IgA, suggesting the necessity for testing all specimens also for IgM. CONCLUSION: Antisperm antibody test results obtained by the IS assay are in agreement with the results obtained with the IBT test. The Immunospheres are monodispersed, color coded, and can be visualized with brightfield microscopy.

Comparison of manual microscopic and computer-assisted methods for analysis of sperm count and motility.

This investigation was conducted to determine which of three methods, manual analysis, and two different commercially available computer-assisted semen analyzers (CASA), was the most reproducible. Semen samples from donors participating in an artificial insemination program (n = 1) and from patients being seen for andrology procedures (n = 12) were acquired at 0.5 h after ejaculation. Each specimen was loaded into one chamber of a 20-microns microcell slide (Conception Technologies, San Diego, CA, USA) and the port was sealed with petroleum jelly to prevent drying of the specimen. The specimens were assessed for sperm count (SC) and motility (MOT) first by manual analysis using an eyepiece reticle and brightfield light microscopy at 400 x total magnification, second using the Hamilton-Thorn 2030 analyzer (Hamilton-Thorn Research, Danvers, MA, USA), and third, using the Cell Trak/S system (CTS; Motion Analysis Corporation, Santa Rosa, CA, USA). Each analysis was repeated five times for each specimen on the same microcell by the same technician. The three methods were compared in terms of means and standard deviations of the SC and MOT over repeated measures-of a specimen using sign tests. The CTS system measured significantly lower sperm counts than the HTM system. MAN was intermediate and not significantly different from either. For MOT, there were no significant differences. Comparison of the standard deviations demonstrated that the three methods were not equally reproducible. For SC, the manual method was significantly less reproducible than the HTM system; the CTS system was intermediate. For MOT, the manual method was less reproducible than either CASA system, both of which were not significantly different from each other. CASA methodology in general provides a more reproducible (less variable) analysis than the manual microscopic method for assessing sperm count and motility.

Differential responses of human sperm to varying concentrations of pentoxyfylline with demonstration of toxicity.

Pentoxyfylline (PF), a methylxanthine derivative, is an inhibitor of the cAMP-phosphodiesterase enzyme, and is known to stimulate the motility of fresh and post-thaw human sperm. The purpose of this study was to examine the effects of different concentrations of PF on motility (MOT), path (curvilinear) velocity (PV), and hyperactivation (HA) of fresh sperm from patients (n = 24) and donors (n = 6) and post-thaw donor sperm (n = 5). For cryopreservation, the donor semen was frozen in liquid nitrogen using test-yolk-glycerol cryopreservative, stored for a minimum of 48 hours, then thawed at room temperature prior to assay. Aliquots of all samples to equal 10 x 10(6)/ml were diluted in 1 ml of the following: medium (human tubal fluid) only (control), or 2.5, 5, 10, or 20 mg/ml PF in medium. Specimens were incubated at 37 degrees C, and all were assessed by computer-assisted motion analysis at 0, 0.5, 1, and 2 hours. The patient specimens were divided into two groups: group 1, mean percent (standard deviation [SD]) MOT < 20% (12.8 +/- 5.8); group 2, mean percent (SD) MOT > 20% (37.8 +/- 14). For fresh donor sperm, 2.5 mg/ml PF significantly stimulated PV and HA at 0, 1, and 2 hours, and MOT at 0, 0.5, and 2 hours. PF at 5 mg/ml resulted in a decreased PV and HA, whereas MOT was decreased by 10 mg/ml. In the < 20% MOT group, 2.5 mg/ml PF significantly stimulated MOT at 0.5, 1, and 2 hours, and HA at 0 and 2 hours. There was no effect on PV.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of low-dose testicular irradiation on sperm count and fertility in patients with testicular seminoma.

The treatment of seminoma with radiation therapy risks transient infertility. We have prospectively followed eight patients with stage I seminoma of the testicle. All patients underwent radical orchiectomy of the affected testis. The mean age of the patients was 32.9 years (range 24-40). Each patient was treated with megavoltage radiation with a 10- or 18-MV linear accelerator. The remaining testicle was shielded using a standard lead enclosure, and the mean testicular dose was 44 cGy (range 20.8-78.2). Semen specimens were delivered to the lab within 30 minutes of ejaculation. All specimens were analyzed using a computer-assisted sperm analyzer. Pretreatment parameters were within normal limits for all but one patient; one patient presented with a borderline normal sperm count at 18 and 22 x 10(6)/ml. Following treatment, there was a decrease in sperm count, detected at 3 months, to < 10 x 10(6)/ml (range 4.4- 8.6 x 10(6)) in all patients except one, who presented with an initial pretreatment count of 189 x 10(6)/ml, which decreased to 58 x 10(6)/ml at 3 months, 32 x 10(6)/ml at 6 months, and rose to 325 x 10(6)/ml by 12 months following treatment. Although the sperm count for this patient (D.L.) was within the normal range, the post-radiation sperm count was less than 20% of the pretreatment count. There was no difference in the motility at 3 months, the mean of which was 51.3%. One patient's (F.C.) wife conceived at 9 months following treatment, one at 12 months (J.R.), and one (J.S.) at 14 months, and all have delivered normal infants.(ABSTRACT TRUNCATED AT 250 WORDS)

Methods of semen preparation for intrauterine insemination and subsequent pregnancy rates.

Semen for insemination, either intrauterine or in vitro, must be prepared to remove seminal plasma products and/or select the healthier population of sperm prior to use. Traditionally, a double wash technique is performed, with or without subsequent swim-up to isolate the motile fraction if necessary. More recently, the use of the SperPrep filtration method has gained acceptance, with the benefits of removal of leukocytes and seminal debris from the specimen as well as enhancement of overall sperm quality. In the current study we compared the traditional double wash method without the swim-up to Sperm-Prep filtration. Intrauterine inseminations (IUI's) were performed in 307 cycles on 148 infertile couples at two different infertility centers in the USA. After complete diagnostic evaluation the couples were offered IUI before proceeding to any other form of assisted reproductive technologies. Semen samples were prepared in human tubal fluid media supplemented with 5% human serum albumin (HSA; location 1) or in Ham's F-10 media supplemented with 3% HSA (location 2), either with the SpermPrep filtration method (ZBL, Inc., Lexington, KY 40523, USA) or the double sperm wash (SW) procedure. Similar sperm numbers were used for the IUI procedure in both treatment groups and locations. The Sperm-Prep method resulted in significantly higher pregnancy rates (PR) than the SW procedure, independent of location. The clinical pregnancy rates per cycle were statistically lower (p < 0.05) in the SW group (20-22% vs. 9-10%). Of significant clinical importance, almost twice as many cycles were required in the SW group to achieve these pregnancies when compared to the SpermPrep group of patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Cryopreservation of human semen. Comparison of cryopreservatives, sources of variability, and prediction of post-thaw survival.

Human semen was cryopreserved using Human Sperm Preservation Medium, TEST-Yolk buffer, or glycerol alone. Sperm characteristics for each specimen were measured before and after freezing to determine which cryopreservative resulted in better cryosurvival and recovery of motile sperm. Sperm frozen in Human Sperm Preservation Medium had a significantly better recovery of all semen parameters (motility, velocity, and recovery) than either TEST-Yolk or glycerol alone. Statistical analyses also were done to examine the variability between and within donor semen specimens. Differences between donors, between specimens, and measurements within donors all contributed to variability of sperm characteristics. Specimen-to-specimen variability for a given donor represented 12% to 47% of the total variability, whereas processing and measurement variability represented 12% to 41%. Donors also varied in the ability of their sperm to tolerate freezing. There was a relationship between motile count after dilution with cryopreservative and post-thaw motile count. This relationship allows the prediction of poor-thaw survival before freezing a specimen.

Pregnancy rates after double versus single insemination with frozen donor semen.

Ninety-nine females (mean age 34 years +/- 1) were seen for therapeutic donor insemination using frozen semen. All patients used over-the-counter urinary ovulation predictor kits, with the insemination scheduled either the day after the positive test (group I; n = 46) or consecutively the day of and the day after the positive test (group II; n = 53). Group I patients underwent a total of 113 cycles of insemination (mean 2.5 cycles/patient). Seven pregnancies were achieved in group I for an overall success of 15.2% and a monthly fecundability of 0.06. Group II patients underwent a total of 100 cycles of therapeutic donor insemination (mean of 1.9 cycles/patient). Twenty-one pregnancies were achieved in this group for an overall success rate of 39.6% and fecundability of 0.21. The estimated cumulative probability of conception (F) for 6 months was 0.32 in group I and 0.78 in group II. These data indicate that the F after therapeutic donor insemination with frozen semen is greater if two consecutive inseminations are performed at midcycle.

Assessment of the viability and acrosome status of fresh and frozen-thawed human spermatozoa using single-wavelength fluorescence microscopy.

The purpose of this study was to use fluorescence microscopy to determine the viability and acrosome status of fresh and frozen-thawed human spermatozoa. Sperm cells were stained with the viability stains Hoechst 33258 (H33258) alone, or propidium iodide (PI) alone, and PI in combination with FITC-conjugated Pisum sativum agglutinin (PSA). The PSA stains the acrosome contents of permeabilized acrosome-intact sperm. Viability by fluorescence microscopy was compared to conventional eosin nigrosin staining. The overall viability using H33258 was not significantly different from that using PI. Therefore, PI was used in combination with PSA for simultaneous measurement of viability and acrosome status at the same excitation wavelength (488 nm). By combining PI and PSA, four subgroups of cells could be detected: group I, PI-neg/PSA-neg--viable, physiologic acrosome reacted (AR); group II, PI-neg/PSA-pos--viable, non-AR; group III, PI-pos/PSA-neg--nonviable, non-AR; group IV, PI-pos/PSA-neg--nonviable, degenerative AR. The postthaw sperm exhibited a significantly greater percent of sperm that were acrosome reacted (both viable and degenerative) (groups I and IV) than the fresh semen. We conclude that frozen-thawed sperm may undergo premature break-down of the acrosome prior to interaction with the oocyte, thus explaining the reduced fertility potential of cryopreserved semen.

Relationship between testicular volume and presence of varicocele. A comparative study.

The varicocele, present in many of the male partners of infertile couples continues to generate controversy, particularly as related to its diagnosis and pathophysiology. The purpose of our study was to determine the relationship between testicular volume and the presence or absence of a varicocele. Testicular volume was determined by the use of an orchiometer; the presence of a varicocele was determined by palpation during a Valsalva maneuver. The patients utilized in the study were those seen in our Andrology and General Urology Clinics; 291 patients with varicoceles and 83 control patients (no evidence of varicocele) were used. The left (mean = 21.4 mL) and right (mean = 23.4 mL) testicular volumes of patients with a varicocele were significantly reduced compared with that of the control group (left, mean = 23.4 mL; right, mean = 26.2 mL; p = 0.0041 and p = 0.002, respectively). The testicular volume corrected on the basis of the body mass (V/m2) also was significantly reduced in the varicocele group compared with controls (left p = 0.007; right p = 0.001). Reduced testicular volume relative to body size may be detected prior to actual demonstration of the stress pattern and the presence of a varicocele, and may be useful in early diagnosis. It is suggested that measurement of testicular volume may be a useful adjunct to routine examination of the infertile male.