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Biomedical subjects

A C Menge

Publications and source records attributed to A C Menge.

At least 37 records · Page 2Linked to original sources

Comparison of intracervical, intrauterine, and intratubal techniques for donor insemination.

OBJECTIVE: To compare the efficacy of intracervical insemination (ICI), intrauterine insemination (IUI), and a combination of intratubal and intrauterine insemination (ITI/IUI) for donor insemination. DESIGN: Prospective randomized clinical trial. SETTING: The University of Michigan donor insemination program. PATIENTS, PARTICIPANTS: Forty-one women undergoing donor insemination with cryopreserved sperm for either isolated male factor or male factor plus ovulatory dysfunction corrected by clomiphene citrate. INTERVENTION: Each patient was randomly assigned to receive each of the three insemination techniques in consecutive cycles until pregnancy occurred or the patient dropped from the study. MAIN OUTCOME MEASURES: Cycle fecundity rates were compared using the chi 2 test, and cumulative pregnancy rates (PRs) determined by life table analysis were compared using a log-rank test. RESULTS: Cycle fecundity rate was significantly higher for IUI (18.3%) than for ICI (3.9%) or ITI/IUI (7.3%). By life table analysis, the cumulative PR for IUI was significantly higher than for ICI, but the PR for ITI/IUI was not. CONCLUSION: For donor insemination with cryopreserved sperm, IUI increases cycle fecundity compared with ICI. The addition of ITI to IUI, however, interferes with the apparent beneficial effect of IUI alone.

Adult↗

Allergic reactions to penicillin during in vitro fertilization and intrauterine insemination.

Two cases are reported in which hypersensitivity reactions occurred after intrauterine placement of spermatozoa or embryos. Because the cells were processed and transferred in antibiotic-containing media, these reactions were suspected to be because of penicillin allergy. One patient had no prior penicillin allergy but was found to be penicillin allergic by skin testing. The other had a history of allergy to oral penicillin. In both cases, the allergy symptoms did not occur during subsequent cycles when antibiotics were removed from the transfer media. These reports suggest that in patients known to be penicillin sensitive, penicillin should not be used during transfer of gametes and embryos for assisted reproductive procedures. In addition, the routine use of antibiotics in these procedures should be scientifically evaluated.

Adult↗

Functional characteristics of sperm obtained by electroejaculation.

Sperm obtained by electroejaculation in 32 anejaculatory men were examined for functional characteristics. Raw specimens showed high sperm counts but motility averaged only 11%. Average viability was 10% for antegrade and 5% for retrograde fractions. Bovine cervical mucus penetration was normal (30 mm. or more in 30 minutes) in only 24% of the electroejaculation samples but it was normal in all of the donor samples tested. Processed sperm motility averaged 30% with 71% forward progression. At 20 hours patient samples retained 46% of the original motility, while donor controls retained 81%. In the hamster egg penetration assay patient sperm penetrated 14% of the oocytes while donor sperm penetrated 40%. Therefore, we identified 4 characteristics of sperm obtained by electroejaculation: 1) low viability, 2) poor survival after overnight incubation, 3) moderately impaired cervical mucus penetration and 4) moderately poor fertilizing capability as measured by the hamster egg penetration assay. Poor sperm survival and impaired function may explain the low pregnancy rates from insemination with electroejaculated sperm.

Adult↗

Fertility of spinal cord injured males: effect of genitourinary infection and bladder management on results of electroejaculation.

Thirty-eight men undergoing electroejaculation (EEJ) procedures for anejaculatory infertility were examined for the presence or absence of infection in urine and semen. In 29 spinal cord injury patients, a high incidence of infected urine and infected semen (41% and 56%, respectively) was seen, in contrast to patients with normal bladder function (0% and 11%). Urinary infection was associated with slightly lower sperm quality and lower pregnancy rates (10% vs 30% in the presence of sterile urine). Semen infection had no effect on sperm counts or pregnancy rates. If intermittent self-catheterization (ISC) was used to empty the neurogenic bladder, slightly better sperm quality was seen, the total failure rate was less, and much better pregnancy rates (44%) resulted than for patients using an alternative bladder management (7%). Antibiotics did not reduce the incidence of urine or semen infection, but did improve sperm counts slightly. Continuous prophylaxis was associated with bacterial resistance to many oral antibiotics and had no advantage over a short course of antibiotics prior to the procedure. Despite the above associations, the sperm quality in our patient population was never normal compared with that of men who ejaculate normally. We conclude that the low sperm quality seen in electroejaculation specimens from spinal cord injured males is not due entirely to infection or to the type of bladder management. Short courses of antibiotics, instead of continuous antibiotic prophylaxis, may be beneficial. Intermittent catheterization is superior to other methods of neurogenic bladder management in maintaining the fertility of spinal cord injured men.

Adult↗

Motility longevity of sperm samples processed for intrauterine insemination.

This study demonstrates that sperm from men with male factor infertility and sperm obtained by electroejaculation have reduced motility longevity when compared with normal specimens. After 24 hours, normal samples lost only 34% of initial motility, whereas male factor patients lost 48%, and electroejaculation patients dropped 66%. Based on these data and previous clinical studies of insemination timing, it is recommended that sperm retrieval and artificial insemination for male factor infertility, especially when electroejaculation is necessary, be performed 24 to 36 hours after urinary detection of the LH surge or as close to the time of ovulation as possible.

Ejaculation↗

Electroejaculation following retroperitoneal lymphadenectomy.

Transrectal electroejaculation was performed in 24 men who were anejaculatory from retroperitoneal lymphadenectomy. Of the men 23 had undergone retroperitoneal lymphadenectomy because of testis cancer. Seminal emission was achieved in all patients. In 21 patients greater than 10 x 10(6) progressively motile sperm with normal morphology were obtained. The average sperm count and motilities obtained were 289 x 10(6) and 18%, respectively, for the antegrade fractions, and 2,051 x 10(6) and 13%, respectively, for the retrograde fractions. Of the 3 azoospermic failures 2 had chemotherapy-induced testicular damage and 1 had carcinoma in situ of the remaining testis. A total of 19 couples underwent artificial insemination with electroejaculated sperm. There were 7 pregnancies achieved in 74 insemination cycles (36.8% of the couples conceived for a 9% cycle fecundity). Routine semen parameters could not predict which couples would be successful in achieving pregnancy. There were 2 first trimester spontaneous abortions. Five healthy children have been born. Electroejaculation is an excellent treatment for anejaculation that persists following retroperitoneal lymph node dissection.

Adult↗

Combined electroejaculation and in vitro fertilization in the evaluation and treatment of anejaculatory infertility.

Seven couples underwent combined electroejaculation and in vitro fertilization for anejaculatory infertility after a failed regimen of electroejaculation and intrauterine insemination. Two pregnancies resulted, one proceeding to a term vaginal delivery and one ending in a 6-week spontaneous abortion. Poor sperm binding and no fertilization were seen in two of the couples. Fertilization of the oocytes but no subsequent pregnancy were seen in the other three couples. Combined in vitro fertilization and gamete intrafallopian transfer was performed in four of the couples. The combination of electroejaculation and in vitro fertilization offers the opportunity to evaluate the female pelvis, observe the sperm-oocyte interaction, and achieve a pregnancy in couples with anejaculatory infertility.

Adult↗

Interrelationships among semen characteristics, antisperm antibodies, and cervical mucus penetration assays in infertile human couples.

Semen characteristics, antisperm antibodies, and cervical mucus penetration studies were analyzed in 754 couples and 95 men undergoing infertility evaluation. The means for the different semen/sperm variables were within ranges published for fertile men. Ages of the men ranged from 22 to 55 years and accounted for a small amount of variation. Sperm counts were lowest in September, December, and January, and highest in April, May, October, and November. Of the sperm characteristics, morphology appeared to be associated with the most other variables. Specimens with more than 50% abnormal sperm forms were overall of significantly poorer quality in terms of sperm counts, motility, forward progression, and ability to penetrate cervical mucus. Antisperm antibodies (agglutinating and immobilizing) were detected in the serum samples of 19.0% of the men, 20.4% of the women, and 32.8% of the couples where one or both partners were positive. Agglutinating antibody titers were significantly correlated between partners. Serum titers of antisperm antibodies were associated with decreased sperm counts, motility, forward progression, and normal forms (immobilizing antibodies). Multiple correlation analysis indicated significant independent effects of sperm concentration, motility, forward progression, and antibodies on sperm-cervical mucus penetration scores of the men. In women, cervical mucus penetration was adversely affected by the presence in the serum of sperm agglutinating antibodies and of immobilizing activity in the cervical mucus.

Adult↗

Predictors of success in electroejaculation of spinal cord injured men.

Rectal probe electroejaculation was attempted in 48 spinal cord injury men and greater than 10 million sperm were obtained in 71%. Patient age and interval since injury had no effect on outcome. The best performance was seen in thoracic paraplegics and in those using intermittent catheterization for bladder management. Indwelling urethral catheters and high pressure reflex voiding had a negative impact on electroejaculation results.

Adult↗

Sperm antibodies in vasectomized men and their effects on fertilization.

Sera (vbs, n = 25) and seminal plasma (vsp, n = 21) from vasectomized men (n = 25) were analyzed for cross-reaction with lithium diiodosalicylate (LIS)-solubilized human sperm extract, protamine, and fertilization antigen (FA-1) with an enzyme-linked immunosorbent assay (ELISA). Among the vbs tested, 44% reacted with human sperm extract, 28% reacted with protamine, and 44% reacted with FA-1 for at least one class of antibodies (IgG, IgA, or IgM). In contrast to the sera, the seminal plasma showed minimal reactions. Neither the vbs nor vsp were found to contain immune complexes, indicating that the antibodies were present in free form. Vasectomized sera that reacted with FA-1 showed a significant (p less than 0.0001) inhibition of human sperm penetration of zona-free hamster ova. The immunoabsorption of FA-1-positive sera with purified FA-1 significantly increased the penetration rates. Affinity-purified human immunoglobulins reactive with FA-1 and not those reactive with protamine reduced sperm penetration rates. Thus, antibodies in vbs reactive with FA-1 are relevant to infertility, causing an inhibition of fertilization. These data will have clinical relevance for diagnosis and treatment of infertility after successful vasovasostomy.

Adult↗

Purification of an acrosomal antigen recognized by a monoclonal antibody and antifertility effects of isoimmune serum.

A highly conserved acrosomal antigen reactive to a monoclonal antibody (HS-63), generated against human sperm, was purified to homogeneity with a combination of conventional procedures and immunoaffinity chromatography using a soluble extract of mouse and rabbit testes. The molecular weight of the purified antigen was 42-50 kD when analysed by sodium dodecylsulphate polyacrylamide gel electrophoresis. The high specificity of the purified antigen to monoclonal antibody HS-63 was shown by indirect immunofluorescent inhibition assay, enzyme-linked immunosorbent assay, Western blot analysis and radioimmunosorbent assay. The purified antigen was used for isoimmunization of mice and rabbits. Following successive immunizations, antisera of high titres were raised and reacted specifically with antigen on the sperm acrosome and in testes of several mammalian species, but not with somatic tissues. These isoimmune sera exhibited strong inhibition on mouse in-vitro fertilization and human sperm penetration of zona-free hamster eggs. The results of this study suggest that the sperm-specific acrosomal antigen reacting with HS-63 could be a good candidate for the development of immunocontraceptive vaccines in humans and in other animals.

Acrosome↗

Characterization of monoclonal antibodies against human sperm antigens by immunoassays including sperm function assays and epitope evaluation.

Fifteen monoclonal antibodies (MAbs) raised against human sperm cells were evaluated for reactions against human sperm by indirect immunofluorescence, immunocytochemistry, agglutination, complement-dependent immobilization, cervical mucus penetration, and hamster egg penetration assays. The MAbs were analyzed for specificity by immunofluorescent reactions with peripheral blood lymphocytes and sperm and classified into three main groups based on regional staining, ie, acrosome, plasma membrane, or tail. One MAb (218) bound to the sperm neck. Three MAbs, (80, 85, and HS-126) were found to react with lymphocytes. Three of five acrosome-reactive MAbs (11, 63, 106), two of five tail-staining MAbs (97, HS-30), and the neck reactor (218) showed significant to highly significant inhibition of sperm penetration of eggs but without significant effects on sperm agglutination, immobilization, or the mucus penetration assay. The three non-specific MAbs gave strong plasma membrane reactions in the agglutination and immobilization assays and also caused highly significant inhibition of sperm penetration of both cervical mucus and zona-free ova. Preliminary analysis of the complementary antigens suggested that epitopes reacting with MAbs 33 (acrosome) and 85 (plasma membrane) were carbohydrate chains on glycoproteins. Three of five MAbs recognizing tail antigen, the neck-staining MAb, and the non-specific MAb (HS-126) appeared to be reactive against glycolipid moieties. Seven of the 12 specific MAbs also reacted in indirect immunofluorescence with mouse and rabbit sperm in patterns similar to those observed with human sperm.

Animals↗

Reduction of fertility in female rabbits and mice actively immunized with a germ cell antigen (GA-1) from the rabbit.

Female rabbits and mice were actively immunized against germ cell antigen (GA-1) of 63 kDa molecular mass isolated from rabbit sperm and testis. There was a significant (P less than 0.05) reduction of fertility in rabbits actively immunized with GA-1 as compared to controls, as seen by the percentage of 9-day implants/corpora lutea ratio (GA-1, 36.3%; controls, 85.7%). In mice, there was again a significant (P less than 0.01) reduction in fertility as seen by mean 7-9 day implants +/- S.D. per mated mouse actively immunized with GA-1 whether through the intraperitoneal route (GA-1, 1.2 +/- 1.6; controls, 8.0 +/- 3.4) or through the subcutaneous/intramuscular route (GA-1, 3.8 +/- 3.4; controls, 10.1 +/- 3.9). The antisera from these actively immunized animals were negative for sperm agglutinating and immobilizing antibodies. In the Western blot enzyme-immunobinding procedure, the antisera showed specific binding to a single protein of 63 kDa. The incidence of fertilization of eggs recovered from rabbits inseminated with anti-GA-1 antibodies-treated sperm was not significantly different from control rabbits. The percentage of fertilized eggs obtained from rabbits inseminated with anti-GA-1 antibodies-treated sperm that reached the blastocyst stage upon in vitro incubation, however, was significantly less than that for embryos obtained from rabbits inseminated with control serum-treated sperm. Incubation of normal fertilized eggs in vitro with the antibodies did not affect development. Neither antiserum nor immune uterine fluid reacted with 4-day blastocysts in the indirect immunofluorescence technique. It is concluded that active immunization with GA-1 results in post-fertilization reduction of fertility in rabbits and mice by inhibiting early embryonic development.

Animals↗

Circulating antisperm antibodies in recurrently aborting women.

One hundred seventy-three women with a history of three or more recurrent consecutive abortions were analyzed for circulating antisperm antibodies with a radiolabeled antiglobulin assay (RAA), a modified enzyme-linked immunosorbent assay (ELISA), a tray agglutination test (TAT), and a sperm immobilization test (SIT). No pregnancies were subsequently gestated to term in women who were antisperm antibody-positive unless they were inoculated with their husband's leukocytes as treatment for an immune basis (not related to antisperm antibodies) for their recurrent abortions. In women with an immune basis for their recurrent abortions, immunization with leukocytes from their male partners increased the ability of these women previously aborting their fetuses to carry their fetuses to term, even if they had positive results in the ELISA, TAT, and SIT; women with positive results in the RAA continued to abort subsequent pregnancies, despite leukocyte immunization. Immunization of antisperm antibody-positive women with their partner's leukocytes did not incite or increase the antisperm antibody titer, with any of the assay techniques.

Abortion, Spontaneous↗

Monoclonal antibodies to human sperm antigens--II.

As part of our continuous effort to elucidate the biochemical and immunological nature of human sperm surface antigens, monoclonal antibodies to human spermatozoa were generated by improved hybridoma techniques. Following immunizations with the membrane fraction of human spermatozoa and cell fusions, hybrid cells were cultured in a semi-solid HAT-selection medium to maximize the number of monoclones recovered. Subcultures were made in liquid phase 7 to 10 days after cell fusions by removing colonies from the initial medium. Based on the results of screening by microplate enzyme-linked immunoassay, 143 of 552 initial clones were found to secrete antibodies to human sperm antigens. More than one-hundred independently derived hybrid cell lines were established. Using indirect immunofluorescent procedures, 62 cell lines were shown to produce antibodies to surface antigens of human spermatozoa. Unique sperm antigens that react with monoclonal antibodies were identified by the SDS gel/protein blot radioimmunobinding method. Sperm agglutinating and immobilizing antibodies were exhibited by 4 and 15 hybrid cell lines, respectively. Fourteen of the monoclonal antibodies also exhibited cross-reactivity with methanol-fixed sperm cells of the rabbit or mouse or both whereas a reaction was not seen with viable sperm of these species. Generation of monoclonal antibodies against a wide spectrum of human sperm antigens should facilitate future investigations regarding immunologic-associated human infertility and fertility control.

Animals↗

Characterization of a membrane antigen from rabbit testis and sperm isolated by using monoclonal antibodies and effect of its antiserum on fertility.

An antigen was isolated from deoxycholate-solubilized rabbit testis and sperm by using an immunosorbent column containing IgG from a monoclonal antibody (8C10.5) that inhibits fertility. Elution was by stepwise increases in pH (8.0, 10.0, and 11.4), with the pH 11.4 fraction after recycling through the column showing a single band at 63 kilodaltons in slab NaDodSO4/PAGE with a silver stain. The antigen molecule was composed of two subunits, which on two-dimensional PAGE showed many spots within the same molecular size range (50-70 kilodaltons) but differing in charge. The antigen isolated either from testis or sperm showed mainly the same spots. The antigen is periodic acid/Schiff positive and contained 21% carbohydrate. An asialo-derivative of the antigen did not change its characteristics on NaDodSO4/PAGE. This glycoprotein resolved into two types of polypeptides, those binding and those not binding to a lens culinaris lectin column; some of the polypeptides appeared common to both fractions. Murine antiserum against the antigen neither agglutinated nor immobilized rabbit sperm but in immunofluorescence reacted with the plasma membrane of viable rabbit sperm as well as with murine and human sperm. Fertilization of female rabbits inseminated with treated sperm was not affected, but, by 9 days, fertility was significantly reduced (21% of controls). The postfertilization antifertility effect was not due to parthenogenic activation or to polyspermy. The antiserum reacted with one specific band in the one- and two-dimensional gel electrophoretic transfer blot procedure and was unaffected by absorption with different somatic tissues.

Animals↗