A novel laboratory procedure to validate American Urological Association guideline on vasectomy success and to diagnose obstructive azoospermia

The objective was to develop a laboratory procedure to validate American Urological Association (AUA) Guideline on vasectomy success when nonmotile spermatozoa are found in the post-vasectomy ejaculate. The neutral α-glucosidase (NAG) an epididymal protein assay modified to determine the activity at 30 and 90 min of incubation from 24 pre- and 47 post-vasectomy ejaculates. The difference between the two points in the relative activity was calculated and if the difference was nonsignificant will confirm vasectomy success. The mean differences in the relative NAG activity were significantly different in pre- and post-vasectomy ejaculates, respectively. The mean differences in the relative NAG activity were similar in post-vasectomy ejaculates with and without nonmotile spermatozoa. No difference in relative NAG activity in post-vasectomy ejaculates between two time points of incubation may be a reliable method to confirm occlusion of the vas deferens. It also validates the recommendation by AUA Guideline on vasectomy success in the presence of few nonmotile spermatozoa.     

Male fertility preservation and cancer treatment

The diagnosis of cancer in men or women leads to prompt evaluation of the extent of the cancer, its treatment, and subsequent prognosis. However, relatively little emphasis is placed on fertility following the completion of therapy. As the effectiveness of cancer treatment has improved, men can enjoy a longer life that is free of cancer. However, chemotherapeutic regimens alone or in combination with radiation therapy frequently result in azoospermia or infertility. This paper reviews available methods to maintain male fertility in patients undergoing chemotherapy or radiation therapy. Certain chemotherapeutic agents that are less likely to cause azoospermia may be incorporated into potentially curative therapies. Hormonal suppression applied early (prior to) and during chemotherapy may protect future male fertility. Alternatively, cryopreservation of sperm enables men to reproduce in the future with the assistance of in vitro fertilization with intracytoplasmic sperm injection. Therefore, oncologists need to discuss male fertility preservation before initiating cancer treatment in reproductive-aged men. The emphasis for future cancer treatment and its research regarding male fertility preservation needs further attention.     

Effect of preincubation of human spermatozoa in milk on penetration of zona-free hamster oocytes: Comparison to TEST yolk

Purpose To investigate the effect of preincubation of human sperm in milk on their ability to penetrate zona-free hamster oocytes, 27 ejaculates were studied. Each ejaculate was divided into two portions and incubated in either milk or TEST yolk (TESTY) for 22–24 hr at 5‡C prior to processing for the sperm penetration assay. Results Spermatozoa preincubated in milk penetrated a significantly (P<0.05) higher percentage of oocytes (mean±SE; 74.4±6.3) than spermatozoa preincubated in TESTY (66.8±6.6). Statistically, no significant differences were noted for penetration index or postincubation sperm motility between the two media. Conclusion Preincubation of sperm in milk therefore appears to enhance their ability to penetrate zona-free hamster oocytes, compared to TESTY preincubation. Since milk is natural, simple, inexpensive, readily available, and can be easily processed, it should be the medium of choice for sperm preincubation.     

Interleukin 1 receptor antagonist gene polymorphisms are not risk factors for recurrent pregnancy loss: evaluation of couples

PROBLEM: The interleukin-1 system has been implicated in pregnancy outcome. Fetal carriage of interleukin-1 receptor antagonist (IL-1Ra) specific alleles has been associated with adverse pregnancy outcomes including spontaneous abortion and pre-term labor. This study was undertaken to compare the frequency of IL-1RN 2 alleles among both male and female partners of couples experiencing recurrent pregnancy loss with that of fertile control couples. METHOD OF STUDY: Buccal swabs were obtained from 42 couples experiencing recurrent pregnancy loss and from 20 fertile control couples. DNA was extracted from the buccal swabs and analyzed for the presence of IL-1RN variable number tandem repeat. RESULTS: No significant differences were found when the frequency of IL-1RN 2 polymorphisms were compared between fertile control couples and couples experiencing recurrent pregnancy loss. Similar results were also obtained when comparing women or men respectively from each group. CONCLUSION: IL-1RN 2 allele is not a risk factor for recurrent pregnancy loss.     

Comparison of thrombophilic gene mutations among patients experiencing recurrent miscarriage and deep vein thrombosis

PROBLEM: Inherited thrombophilia has been shown to be a risk factor for cardiovascular disease including deep venous thrombosis as well as reproductive disorders including recurrent pregnancy loss. We have previously reported three out of the 10 thrombophilic mutations studied, plasminogen activator inhibitor-1 (PAI-1) 4G/5G, factor XIII V34L, and homozygous MTHFR C667T, correlated significantly with recurrent pregnancy loss compared with controls. This study was undertaken to compare the frequencies of nine inherited thrombophilias among women with a history of recurrent pregnancy loss with individuals experiencing deep venous thrombosis and fertile controls. METHOD OF STUDY: Six hundred thirty-four participants including 550 women with a history of recurrent pregnancy loss, 43 individuals with deep vein thrombosis and 41 fertile women without a history of recurrent miscarriage. All participants had buccal swabs taken for DNA analyses of nine gene polymorphisms including factor V G1691A, factor V H1299R (R2), factor II Prothrombin G20210A, factor XIII V34L, beta-fibrinogen -455G>A, PAI-1 4G/5G, human platelet antigen 1 a/b (L33P), MTHFR C677T, MTHFR A1298C. Frequencies of thrombophilic gene polymorphisms were compared among the three populations studied. RESULTS: Individuals with a history of DVT had a significantly higher frequency of all of the polymorphisms studied compared with women experiencing a history of recurrent pregnancy loss and the fertile controls. The frequencies of mutations for V34L and PAI-1 4G/5G were significantly increased among women experiencing recurrent pregnancy loss compared with controls. The most prevalent polymorphisms were factor XIII V34L and PAI-1 4G/4G for both individuals with a history of deep vein thrombosis and recurrent pregnancy loss compared with controls. CONCLUSION: Screening for risk factors for inherited thrombophilia with only polymorphisms for factor V von Leiden, factor II prothrombin and MTHFR may be missing the more prevalent identifiers of jeopardy.     

Anterior shoulder dislocation with axillary artery and nerve injury

We report a rare case of left axillary artery injury associated with anterior dislocation of the left shoulder in a 25 yrs old male as a result of a road traffic accident. The shoulder dislocation was reduced. A left upper limb angiogram showed an obstructed left axillary artery. The obstructed segment was surgically reconstructed with a Dacron graft. Six months post operation in follow up, he was found to have good left shoulder function and no neurovascular deficit. This is an injury that could have been easily missed without a simple clinical examination.     

Cystic fibrosis mutation screening in healthy men with reduced sperm quality

The majority of men with cystic fibrosis (CF) are infertiledue to a bilateral congenital absence of the vas deferens (CBAVD).However, clinically affected CF patients present a spectrumof genital phenotypes ranging from normal fertility to severelyimpaired spermatogenesis and CBAVD. Recently, it has becomeapparent that CF can manifest itself as isolated CBAVD in theabsence of other clinical symptoms. The present study was undertakento test the possible involvement of the CF gene in the aetiologyof male infertility other than CBAVD. Semen specimens from 127unrelated healthy males with various diagnoses of reduced spermquality were screened for a panel of 13 mutations in the cysticfibrosis transmembrane conductance regulator (CFTR) gene. Fourteenof 80 (17.5%) healthy men with infertility due to reduced spermquality and 3 of 21 (143%) men with azoospermia had at leastone CF mutation (one azoospermic male was a compound hetero-zygote).The frequency of mutations in our sample of infertile maleswas significantly higher than the expected CF carrier frequencyin the local population (P = 0.00139). No mutations were foundin a control group of 26 individuals with normal semen parameters.This increased frequency of CF mutations in healthy men withreduced sperm quality and in men with azoospermia without CBAVDsuggests that the CFTR protein may be involved in the processof spermatogenesis or sperm maturation apart from playing acritical role in the development of the epididymal glands andthe vas deferens.     

Variation in Antisperm Antibody Results Using Different Assays

PROBLEM: The detection of various types of antisperm antibodies (ASA) in the serum varies among different assays. This variation may influence the diagnosis and management of infertile couples who are tested for such immunologic factors. This prospective study was conducted to determine the variation in the results of ASA as measured by the sperm immobilization (SI), sperm agglutination (SA), and the indirect immunobead (IB) assays. METHOD: The sera of 79 patients that tested positive for ASA by at least one of the assays listed above were concurrently tested with all three assays. RESULTS: Using an individual ASA assay, 66 (84%), 26 (33%) or 36 (46%) of sera tested positive by the SA, SI, or IB assays, respectively. However, using a combination of assays, 67 (85%), 78 (99%) or 40 (51%) of sera tested positive using either the SA+SI, SA+IB or SI+IB assays, respectively. CONCLUSION: These findings suggest that the utilization of different assays to detect ASA may detect sera that are positive for ASA with more reliability than single assay testing.     

The production of interleukin-1β from human fetal membranes is not obligatory for increased prostaglandin output

Bacterial endotoxin increased the expression of mRNA (maximal after 4 hr) for interleukin-1beta (IL-1beta) and the release of mature protein from intact human fetal membranes. In contrast, the change in expression of mRNA for type 2 cyclo-oxygenase (COX-2) was biphasic, with peaks after 0.5-1 hr and after 8 hr of culture. An antibody to IL-1beta was without effect after 4 hr of culture, inhibited endotoxin-stimulated prostaglandin E2 (PGE2) production after 8 hr of culture, and caused a parallel decrease in the expression of mRNA for COX-2. We conclude that endotoxin induced the expression of COX-2 through IL-1beta-independent and IL-1beta-dependent mechanisms, and these differences are time dependent. Corticotrophin-releasing hormone (CRH) or platelet-activating factor (PAF) also increased the expression of mRNA for IL-1beta and the release of IL-1beta from some, but not all, fetal membranes. The antibody to IL-1beta did not affect CRH-stimulated or PAF-stimulated PGE2 production or COX-2 expression. We conclude that CRH and PAF can induce the expression of IL-1beta, but this is not obligatory for increased PGE2 release, and the effect of these stimuli on COX-2 expression is a direct, IL-1beta-independent effect.