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Studies on the immunogenicity of protamines in humans and experimental animals by means of a micro-complement fixation test.

A complement fixation study with human, monkey and rabbit sera, using purified sperm nuclear basic proteins as antigens, led to the following conclusions. (1) Protamines, the sperm-specific basic nuclear proteins, may be immunogenic in mammalians. (2) Antibodies detected in the indirect immunofluorescence test on human swollen sperm heads in sera from infertile and vasectomized men, are directed primarily against human protamines. (3) The results obtained suggested that differences in the immunization site and/or in the configuration of the immunizing protamine, may lead to the formation of antibodies directed against different antigenic determinants. Autoimmunity to protamines, following vasectomy or in infertile men, is accompanied by the formation of antibodies cross-reacting with common antigenic determinants present in protamines of other species. Induction of immunity to protamines by means of immunization with protamines-RNA complexes (in rabbits), or protamine-insulin complexes (in humans), leads to the formation of antibodies reacting more specifically with the immunizing protamine, showing only slight cross-reaction with other protamines. (4) The histone-like fraction present in mature human spermatozoa is composed mainly of histone fraction H2B.

Animals↗

[The filtration capacity of the cervix uteri (in-vitro studies) (author's transl)].

In-vitro experiments had been undertaken in order to clarify sperm selection by cervical mucus. Sperm samples, preferably with a high rate of abnormal cells, were obtained from fertile and infertile men for 35 penetration tests. Cervical mucus was collected from fertile women during praeovulatory phase. The penetration tests resulted in a statistically high amount of normal sperm cells which had migrated through cervical mucus as compared to abnormal cells. The significance of this filtration capacity of the cervical mucus will be discussed.

Cervix Mucus↗

The necessity for bilateral biopsy in oligo- and azospermia.

The 384 testicles from 192 infertile men were examined by bilateral biopsy. The main reason of this investigation was to see whether differences in the varying degrees of damage to the seminal epithelium existed in both testicles. Depending on the damage, the material was divided into five groups. The conclusion is, that the greater the degree of damage, the greater are the differences in the findings in both testicles. The left testis was more often and more strongly damaged than the right.

Adult↗

Sperm surface proteins after capacitation. Expression of vitronectin on the spermatozoan head and laminin on the sperm tail.

Several integrins recognize as ligands proteins containing the Arg-Gly-Asp (RGD) sequence, such as fibronectin, vitronectin, and laminin. It has been previously demonstrated that oligopeptides containing the RGD sequence competitively inhibit both the adhesion of hamster and human sperm to zona-free hamster eggs and their subsequent penetration. In addition, the appearance of fibronectin on the surface of living human spermatozoa after capacitation has been demonstrated. In this work, it is shown that spermatozoa incubated overnight under capacitating conditions, but not fresh spermatozoa, also display the RGD-containing proteins vitronectin and laminin. Whereas the expression of fibronectin does not appear to be localized to any specific region of the sperm surface, laminin is present solely on the sperm tail, and vitronectin was detected mostly as an equatorial band on the sperm head. The percent of capacitated spermatozoa within each ejaculate reacting with antivitronectin antibodies (51% to 94%) was similar to that observed with antifibronectin antibodies (72% to 100%) in a series of fertile donors, and in a series of infertile men (7% to 98% for vitronectin versus 5% to 100% for fibronectin). In contrast, the percent of spermatozoa displaying laminin was lower, ranging from 2% to 42% for fertile donors and from 5% to 34% for infertile donors, and was unrelated to the expression of fibronectin or vitronectin. The time of appearance of both fibronectin and vitronectin when spermatozoa were incubated under capacitating conditions varied for different sperm donors, suggesting a difference in the process of their expression between different men. The specificity of antivitronectin antibody binding to human spermatozoa was demonstrated by competitive inhibition with purified human vitronectin. That there was no immunologic reaction between the antivitronectin antibodies used and fibronectin was demonstrated both by the failure of free fibronectin to inhibit antivitronectin antibody binding to spermatozoa, and by Dot blot analysis. A partial cross-reaction of the polyclonal antivitronectin antibody with fibronectin was shown by Western blot analysis, but this phenomenon was not present when the monoclonal antivitronectin antibody was used. In addition, both fibronectin and vitronectin could be extracted from capacitated spermatozoa solubilized in Chaps buffer, as shown by Dot blot and Western blot analysis. These observations suggest that vitronectin and fibronectin expressed on the surface of capacitated human spermatozoa could act as a ligand for specific receptors on the egg, and play a role in sperm-oolemmal adhesion.

Blotting, Western↗

Testicular biopsy in male infertility.

A total of 134 testicular biopsies from infertile men were studied. The potentially treatable causes of infertility viz. Normal spermatogenesis with post-testicular obstruction, maturation arrest and hypospermatogenesis together constituted about 54 percent of all cases. The incurable causes viz., germ cell aplasia, tubular fibrosis and Klinefelter's syndrome together constituted about 42 percent of all cases. Testicular biopsy in male infertility is a very useful procedure since it gives a lot of information on the diagnosis and helps in planning treatment.

Biopsy↗

[Dynamic tests with GnRH, L-dopa, clomiphene, cyclophenyl, HCG and HMG in 98 cases of dysspermia due to a variety of causes].

Radioimmunological methods have been used to assay the plasma concentrations of gonadotropins, androgens (expressed as the sum of testosterone and dihydrotestosterone) and 17beta-oestradiol in 98 patients suffering from dysspermia of variable etiology, after stimulation with GnRH, Clomiphene, L-Dopa, Cyclophenyl, HCG in various dosage schemes, and HMG. Results show L-Dopa and HMG tests to be of little value in assessing the endocrine function in infertile men. They also confirm, in keeping with data found in the literature the usefulness of dynamic stimulation with GnRH, Clomiphene, and HCG for purposes of andrologic diagnosis. The GnRH proved useful for the distinction between primary and secondary hypogonadotropism as well as for assessing the therapeutic efficacy of releasing factors. The functional efficacy of the endocrine axis which is often impaired in patients with dysspermia can be evaluated by the Clomiphene test. In the small number of cases in which it was applied, Cyclophenyl brought about changes in gonadotropin concentration which require further study. The HCG tests, to be performed in patients with lower than normal testosterone plasma lavels, permits the discrimination between primary and secondary Leydig cell deficiencies as well as the assessment of interstitial tissue sensitivity to chorionic gonadotropin stimulation. Of the three dosage schemes (5000 IU, 7500 IU "step up", 10.000 IU) the last one is most suitable for clinical use.

Adult↗

Immunology and male infertility.

This article has attempted to review evidence that suggests that immune factors may be operative in a small but significant number of infertile men. Although our current understanding of the possible processes by which autosensitization to previously sequestered reproductive antigens occurs is incomplete, there are laboratory assays presently available that give an indication, but do not prove, that immune factors may be contributing to the infertile state. Continued research is needed to develop new assays and more purified sperm antigens, which might enhance our knowledge of the underlying immunoreproductive changes. Until these are available, the following procedures should be considered when investigating a patient with infertility of suspected immune origin. The patient's history should be taken, and a physical examination should be performed. A complete blood count, urinalysis, and complete semen analysis and culture should be taken. Next, in vivo cervical tests (Sims-Huhner) are performed, followed by sperm antibody assessment (serum, semen) and perhaps in vitro cervical mucus sperm assays (especially the crossed hostility test). After the tests have been completed, the following possibe treatments exist: 1. Treatment of underlying infections 2. Correction of obstructions 3. Corticosteroid (or testosterone?) therapy 4. Washed sperm insemination 5. First portion of fresh ejaculate insemination 6. Artificial insemination with homologous donor 7. Adoption.

Agglutination Tests↗

Immunity and male infertility.

This article has attempted to review evidence which suggests that immune factors may be operative in a small but significant number of infertile men. Although our current understanding of the possible processes by which autosensitization to previously sequestered reproductive antigens occurs is incomplete, there are laboratory assays presently available that indicate, but do not prove, that immune factors may be contributing to the infertile state. Continued research is needed to develop new assays and materials (especially purified sperm antigens) which might enhance our knowledge of the underlying immunoreproductive changes. Until these are available the outline below might be considered when investigating a patient with infertility of suspected immune origin.

Animals↗

Carcinoma-in-situ of testis in testicular feminization syndrome.

A light microscopical study of an excised testis from a 14 1/2-year-old girl with incomplete type of testicular feminization syndrome revealed a carcinoma-in-situ pattern in a part of the gonad. An identical histological pattern has repeatedly been demonstrated in infertile men who developed gross testicular germ cell tumours. It is suggested that germ cell carcinomas in patients with testicular feminization syndrome may be preceded by this characteristic intratubular germ cell abnormality.

Adolescent↗

On round-headed human spermatozoa.

In an infertile man and his only, likewise infertile, brother the sperm contained only round-headed spermatozoa. Electron microscopic examination of embedded ejaculate revealed a malformation, viz. absence of acrosomes in all the spermatozoa with consequent lack of normal development of the heads of the spermatozoa. Acrosomeless spermatozoa were also found in the testicular tubuli. No disturbance of the endocrine functions could be demonstrated. The blood cells were of normal male karyotype. Investigation of meiosis, on the other hand, showed rudimentary second division. Two other unrelated infertile men showed the same isolated disturbance in their spermiogram. Treatment with chlomiphene had no effect on the structure of the spermatozoa or on the infertility. The rare disorder, which seems to be a special syndrome, the first hitherto distinguishable among the various types of teratospermia, and its probable background are discussed.

Acrosome↗

Leydig cell density and function and their relation to gonadotropins in infertile oligospermic men with varicocele.

Circulating gonadotropin and testosterone levels in infertile men with varicocele and sperm counts of less than 10(7)/ml were correlated with testicular Leydig cell density and in vitro testosterone synthesis. A significant correlation was found between Leydig cell density and both plasma testosterone levels and in vitro testosterone synthesis. Luteinizing hormone (LH) levels paralleled the in vitro testosterone synthesis. Although in the majority of patients, in vitro testosterone synthesis and Leydig cell density were low, plasma testosterone and LH levels were within the normal range. The only abnormal circulating hormonal level was high follicle-stimulating hormone in the most severely oligospermic patients. No difference were found between the right and left testes with regard to in vitro testosterone synthesis or Leydig cell density, and no correlation with patient age or site or degree of varicocele could be demonstrated. These findings suggest that Leydig cell dysfunction was the mechanism responsible for the oligospermia and infertility of these patients.

Adult↗

Improvement of semen and pregnancy rate after ligation and division of the internal spermatic vein: fact or fiction?

Male partners with left-sided varicoceles of 96 infertile couples were studied. Fifty-one patients were submitted to ligation of the testicular veins and 45 individuals were randomised as controls. During an observation period of 53 months (range 36 to 74 months) we found no statiscally significant improvement in the semen crude variables, the morphology or the progressive motility in the series of men submitted to surgery. The pregnancy rate was lower in those who had an excision of varicocelle.

Adult↗

Value of a testicular biopsy rating for prognosis in oligozoospermia.

Testicular biopsies of 142 oligozoospermic men were used to obtain a testicular biopsy score count. These scores were clearly related to the chance of fertility, in contra-distinction to data on hormone concentrations and from analysis of semen. In 36 patients with a score of 9-10 there were 15 pregnancies; in 59 patients with a score of 8-9, 12; and in 47 patients with scores below 8 there was only one successful pregnancy, though this last group also contained an additional three patients whose wives' pregnancies ended in miscarriages. Rating of testicular biopsies is more useful for proper evaluation of oligozoospermic patients than are data on hormone concentrations and from semen analysis. Proper evaluation of new treatments should be based on trials where experimental and control groups are matched on these ratings.

Biopsy↗

[Examination of male sterility].

There is a description of the main principles for the exploration of male fertility and the evaluation of the results thus obtained. In spermiological investigation the importance of an objective evaluation of sperm motility is emphasized. There is a well-arranged outline of the lower standard limits and the average spermiological values in ejaculates of normal fertile men as well as the nomenclature, both the one used in Prague Institute of Sexology and the suggestion of the international nomenclature given by the Andrology Club.

Humans↗