Detection of high-risk HPV DNA in semen and its association with the quality of semen.

The effects of seminal high-risk human papillomavirus (HPV) DNA were assessed on the quality of semen. Semen samples of 65 men participating in the ongoing Finnish HPV Family Study were collected. Semen analyses were done by the guidelines of the Nordic Association for Andrology. HPV DNA was detected by nested polymerase chain reaction and confirmed by Southern blot hybridization for high-risk types. Altogether, 10/65 men (15.4%) had high-risk HPV DNA positive semen sample. Seminal high-risk HPV DNA did not affect semen volume, sperm concentration, motility and vitality of spermatozoa. However, semen pH was borderline lower in HPV DNA positive than negative samples (7.4 vs 7.5). Neither oligo- nor asthenozoospermia was associated with seminal HPV DNA. In conclusion, seminal high-risk HPV DNA was detected in 15% of men. It did not affect the semen analysis, except semen pH by borderline significance. Sperm donors have not been tested for HPV infections, sperm washing does not seem to eliminate the risk of HPV transmission and the consequences of HPV in the semen are at present unknown.

Sympathetic auto-immune orchitis.

There is increasing evidence that, besides the immunization to sperm antigens and other immunological disorders, spontaneous autoimmune orchitis may also be an aetiological factor in male reproductive failure. The present paper describes a case study, where a surgical operation for an inguinal hernia caused an orchitis first in the ipsilateral, and few weeks later also in the contralateral testis and which finally led to an atrophy of both testicles. The time-course, patient's symptoms and especially the final histological picture all indicate that this process had an immunological basis, and therefore it is proposed that it should be called 'auto-immune sympathetic orchitis'.

Successful treatment of infertility due to retrograde ejaculation by instillation of serum-containing medium into the bladder. A case report.

The treatment of infertility caused by retrograde ejaculation has often been ineffective. The unphysiological composition of the urine is the main reason complicating the recovery of motile sperm. We describe a case of retrograde ejaculation caused by a congenital defect of the internal sphinchter muscle of the urinary bladder. In this case, motile sperm were only recovered if the bladder had been filled previously with artificial culture medium (Ham's F-10) supplemented with the patient's own serum. Two pregnancies followed intrauterine insemination with sperm recovered in this way.

Interaction of male and female sex hormones in cultured rat prostate.

The organ culture of the rat ventral prostate was chosen as a model to determine whether any of the estrogen effects in vivo on the prostate are direct and expressed at the hormone concentrations normally found in the male. During 2 weeks of culture, estradiol at the high concentration of 10(-5) M blocked the androgenic activation of [3H]thymidine incorporation into DNA. The inhibition was localized in epithelium. Protein content of testosterone-treated explants and the accumulation of prostatein in the medium were considerably decreased, indicating inhibition of secretion. Antiandrogenic effects were not seen in morphology of estrogen-treated explants. The lower concentrations (from 10(-9) M to 10(-6) M) of estradiol increased the volume density of epithelium from day 7 onwards. The height of epithelium was concomitantly increased. The volume density of epithelium as well as the percentage of acini with metaplastic changes were significantly increased. These epithelial changes were less pronounced in the presence of androgen, suggesting that physiological concentrations of androgen prevent the expression of estrogen action in the morphology of the prostate. A change in staining with peanut (PNA)- and wheat germ agglutinin (WGA)-lectins indicated defective secretory capacity in metaplastic epithelium. In spite of the increased protein content in the explants, no constant pattern of the changes in prostatein accumulation could be recorded. Although the concentrations of estrogen required to induce squamous metaplasia were still unphysiological, the occurrence of this abnormal differentiation of the prostatic epithelium suggests that the cooperative action of estrogen is involved in androgen-dependent normal epithelial growth and possibly also in promoting growth of prostatic neoplasia.

Expression of acrosin during mouse spermatogenesis: a biochemical and immunocytochemical analysis by a monoclonal antibody C 11 H.

A monoclonal antibody C 11 H was produced against human acrosomal antigen. It also cross-reacted with acrosomes of the boar and the mouse. In boar spermatozoa the antibody reacted, in immunoblotting analysis, with polypeptides of Mr 55,000 and 53,000. The proteolytic activity was detected by zymographic casein overlay assay. Partially purified boar sperm acrosin was bound by the C 11 H affinity column, and acrosin activity was detected in the eluate. These experiments indicate that C 11 H antibody recognizes sperm acrosin. The acrosin expression during spermatogenesis was studied with C 11 H antibody, using mouse testis as a model. Immunocytochemical analysis revealed that step 9 spermatids were the first cells to react with C 11 H antibody. During step 14, the perinuclear pattern of C 11 H-binding disintegrated into small particles around the spermatid nuclei for the period of close association between spermatid bundles and Sertoli cells. During late step 15, the antigen became located at the site in the acrosome typical for step 16 spermatids and spermatozoa. These results indicate that monoclonal C 11 H antibody recognizes acrosin that is first expressed in haploid cells coincident with the onset of nuclear elongation and cessation of RNA transcription. The changes in the distribution pattern suggest that acrosin may be modified by Sertoli cells. In addition to studies on acrosin, this antibody may be useful in investigations of transcription and translation and their regulation during spermatogenesis in general.

Testicular plasminogen activators during postnatal development in the rat.

In the seminiferous epithelium, Sertoli cells secrete plasminogen activator (PA) under regulation of follicle stimulating hormone, cyclic AMP and neighbouring spermatogenic cells. Recent observations suggest that preleptotene spermatocytes upon their release from the basement membrane of the seminiferous tubule are important regulators of PA secretion. To study further the role of PA's in the seminiferous tubules, we have analyzed the endogenous levels and secretion rates of PA at various ages during postnatal development, and performed biochemical analyses of the types of PA in the testis and spent media from seminiferous tubular cultures. Cyclic secretion of PA started at the age of 28 days, and from 40 days onwards, the high secretion rates were localized in stages VII and VIII of the cycle of the seminiferous epithelium. The secreted PA is most obviously of the urokinase type; both urokinase-type and tissue-type PA-like activities were found in seminiferous tubular homogenates. The increase in testicular PA levels concomitant to the onset of meiosis in the epithelium was due to the urokinase-type PA-like activity.

Laminin and type IV collagen in the human testis.

Specimens of normal human testis and biopsies from testes with Sertoli-cell-only syndrome in which the seminiferous tubules had a remarkably thickened lamina propria, were investigated immunohistochemically using specific antibodies against human laminin and human type IV collagen. In the normal testis, both laminin and type IV collagen were localized to the epithelial basement membranes and the peritubular cell layers. In addition, laminin was found in the Sertoli cells. In the pathological testis, structures representing invaginations of the tubular basement membrane were positive for both laminin and type IV collagen. The presence of laminin and type IV collagen in the myoid cell layers, and laminin in the Sertoli cells from both normal and pathological testis and its indication for the secretion of these substances by the myoid and Sertoli cells is discussed.

Cellular regulation of plasminogen activator secretion during spermatogenesis.

The secretion of plasminogen activator (PA) has been found to be highly stage-specific during rat spermatogenesis. It is maximal in Stages VII and VIII of the cycle. At these stages, seminiferous tubules contain primitive type A1 spermatogonia, preleptotene and midpachytene primary spermatocytes, round and maturation-phase spermatids and Sertoli cells. The last cell types are the most likely sources of PA. To investigate which cell type might be involved in the regulation of PA secretion, we have sequentially isolated 1-mm segments of rat seminiferous tubules from Stages VI-IX with transillumination-assisted microdissection and measured PA secretion using 125I-labeled fibrinogen as substrate. In another experiment, spermatogenia were killed by 300 rads of x-rays and PA secretion was analyzed during the absence of desired germ cell classes. The results support the idea that upon their detachment from the basal lamina, preleptotene-stage primary spermatocytes have the major stimulatory action on the PA secretion of the seminiferous tubules. Other phenomena such as spermiation, phagocytosis of residual bodies or opening up of the Sertoli cell junctions seem to influence PA secretion to a lesser extent.

A comparison of eight techniques for the evaluation of the auto-immune response to spermatozoa after vasectomy.

The tray agglutination tests (TAT), gelatin agglutination test (GAT), side agglutination test (SAT), tube-slide agglutination test (TSAT), sperm immobilization test (SIT), ATP-release cytotoxicity test (ARCT), indirect immunofluorescence technique (IFT) on methanol-fixed, intact spermatozoa, and a lymphocyte transformation test (LTT) were compared using a maximum of 329 blood samples taken from 47 men before and after vasectomy. The TAT, GAT, TSAT, SIT and ARCT discriminated between the pre- and post-vasectomy samples, and the sensitivity for sperm antibodies decreased in that order. The activity in the IFT and the LTT did not change significantly after vasectomy. In the TAT the mode of agglutination varied with serum dilution; the results for the 1:4 dilution showed the best agreement with the SAT results. Almost all TAT activity was detected by a combination of GAT and TSAT. Sperm agglutinins were present in all serum samples positive in the two complement-dependent tests, SIT and ARCT. If improved in sensitivity, the ARCT, which lacks the subjective elements of microscopy, might be suitable for the screening of male sera in clinical work. For the present, we recommend the TAT.

A new method for measurement of cytotoxic antibodies to human spermatozoa.

The energy producing reactions of spermatozoa will cease after the spermatozoa have been damaged by a complement-mediated antibody reaction and, consequently, their ATP content falls rapidly. The finding was applied and used for determination of the cytotoxic reaction caused by sperm antibodies. This reaction could be recorded by comparing the difference in ATP content between damaged and control spermatozoa after 2 hr incubation. A serum dilution that still reduced the ATP content by 50% or more was regarded to be cytotoxic. Only those sera, collected from infertile patients, with an agglutinating titer of at lease 1:64 and an immobilizing titer of 1:4 were cytotoxic to spermatozoa. The method proved to be both simple and more sensitive than the supraviatal staining technique as a cytotoxicity test.

Improvement of spermatozoal motility characteristics.

In this study were collected 34 normal semen samples, and samples from 8 men showing prostatic hypofunction of unknown origin and from 5 men with agglutinating autoantibodies in their sera. Among the various biological solutions tested, Tyrode's solution, with or without glucose, bovine serum albumin, and sperm-Ringer solution containing fructose best maintained the motility of normal spermatozoa, which was nearly the same as with semen samples. The ATP content of the spermatozoa was highest in the Tyrode's solution. When seminal plasma of the samples with low acid phosphatase and poor sperm motility was replaced with normal donor seminal plasma, the motility was increased almost to the normal level. All other biological solutions tested were weaker in this respect. 7.2 mmol/l caffeine had neither clear improvement on the motility of normally motile sperm cells nor on the spermatozoal ATP content. After cryopreservation and thawing, the samples containing caffeine showed the best motility, which was, however, surprisingly low. Spermatozoa coated by autoantibodies showed very abnormal cervical mucus penetration. After washing with Tyrode's solution the penetration improved, but did not reach normal penetration level.