Clinical Andrology Research Articles

Semen quality and human fertility: a prospective study with healthy couples.

Measures of semen quality are used as surrogate measures of male fertility in clinical andrology, reproductive toxicology, epidemiology, and risk assessment. However, only limited data are available to relate those measures to fertility. This prospective study with 210 reproductive-age couples was conducted to provide information on the value of semen quality measures for predicting human male fertility potential and for development of models to estimate the effects of changes in semen quality on fertility in a given population for risk assessment. Couples without known risk factors for infertility and who had discontinued contraception to have a child were accepted. The study followed each couple for up to 12 menstrual cycles while they attempted to conceive and evaluated semen quality measures from multiple ejaculates per man with known abstinence intervals. For each cycle, the day of ovulation was predicted, and the couple was advised to have intercourse multiple times on that day and on the days around it. Among the demographic variables assessed, parity, contraception status prior to entering the study, male education level, and male smoking were associated significantly with 12-cycle pregnancy rate. Several semen quality measures also were associated significantly with pregnancy rate, with percentage morphologically normal sperm by strict criteria and measures involving total number of sperm showing particularly strong associations. Localized regression-smoothing plots of semen quality data against proportion of couples pregnant suggested levels below which fertility declines for several semen quality measures. These results have applications in both clinical andrology and in assessment of risk to male fecundity from environmental or pharmaceutical exposures. In particular, they contribute information on behavior of fertility with varying semen quality and can allow development of models to predict effects on fertility in populations from decrements in semen quality.

Non-surgical sperm recovery: part I.

For years, clinical andrology has been a discipline full of tradition and history. In the beginning clinical andrology was mainly the province of endocrinologists who were initiated in the secrets of the hypothalamo–pituitary–gonadal axis of men. Their treatments modulated this endocrinological axis in an attempt to improve spermatogenesis. Then urological surgeons introduced reconstructive surgical techniques. Since the 1980s, gynaecologists and clinical embryologists entered the field. They introduced techniques of assisted reproduction which were more successful than the common empirical treatments (O’Donovan et al., 1993). However, the major breakthrough came only with the introduction of intracytoplasmic sperm injection (ICSI) (Palermo et al., 1992). When injected into a metaphase II oocyte, a spermatozoon was found capable of activating this oocyte without further manipulations. The ease by which human oocytes are fertilized by spermatozoa, even if these spermatozoa are immature, has led to a widespread use of this technique of assisted reproduction. Because of this success, the clinical work-up of the patient with poor sperm quality is becoming more and more endangered (Devroey et al., 1998). A typical attitude, common among many fertility ‘specialists’ nowadays, can be summarized as follows: ‘catch the sperm and inject!’. However, without a proper clinical work-up and approach, an azoospermic man with a Kallman syndrome may one day be scheduled to undergo a testicular biopsy to retrieve sperm for ICSI! A great deal of apprehension exists concerning the safety of ICSI (Cummins and Jequier, 1995). It is the role of the clinical andrologist not only to try to elucidate the pathophysiological mechanisms behind a man’s reproductive failure, but also to formulate the most cost-effective and safest treatment. Only a rigorous clinical work-up may ensure that techniques of assisted reproduction such as ICSI are applied only when strictly indicated and when no alternative treatments are present. The introduction of surgical sperm recovery for ICSI has given the possibility to many sterile men to father children which are genetically their own. However, because of the simplicity of these surgical sperm recovery techniques, there is a tendency to propose ICSI with testicular sperm as a first-line treatment in many patients with ejaculatory disturbances as well. However, the long-term ramifications of using these immature spermatozoa for ICSI (Aslam et al., 1998), means that non-surgical methods are preferable in these patients in order to recover mature spermatozoa. These non-surgical methods are often more cost-effective and although they may not preclude the need for ICSI, they may offer the opportunity to use more mature spermatozoa. There are also other advantages. In patients with anejaculation because of spinal cord injury, ejaculated spermatozoa may be obtained by vibrostimulation or electro-ejaculation. Even if the quality of the semen obtained may be poor and often ICSI still has to be performed, the non-invasive character of these techniques represents a real advantage for these patients. Because of their neuro-vascular impairment, procedures such a testicular biopsy should be avoided as much as possible in order to prevent surgery-related complications. It will become obvious to the reader of this mini-symposium, published in two parts, that nonsurgical sperm recovery is a far neglected, but nevertheless very important area in the field that should be familiar to every clinician involved with clinical andrology.

Localisation of enzymes in live spermatozoa by CellProbe reagents (preliminary results).

As a new approach, various synthetic fluorogenic substrates, the CellProbe reagents, were applied to examine the topography of their cleavage in vital human spermatozoa. These substrates are able to enter the cells without requiring previous cell permeabilization and can produce a fluorescent dye after cleavage, depending on enzyme activity. Vital spermatozoa from samples with normal spermiogram parameters showed fluorescence in different areas and intensity after incubation with a variety of substrates for aminopeptidase A, peroxides, subtilisin, dipeptidylpeptidase IV (DPP IV), cathepsin D, glucosidase and glucuronidase, but not with the substrate for galactosidase. Fluorescence was mainly located in the acrosomal cap (substrates for DPP IV, subtilisin, cathepsin D, glucosidase and glucuronidase) in the middle piece and head (substrates for peroxides, glucosidase), in the sperm head (substrates for aminopeptidase A) and occasionally in the tail (substrate for glucosidase). The substrate for subtilisin may play a role in andrology, because subtilisin is a serine protease like acrosin. This substrate may possibly be used to determine the acrosin activity in vital spermatozoa. The CellProbe reagents for fluorescence cytoenzymology may serve advanced methods in both clinical andrology and spermiological research, presuming that the characteristics and qualities of the synthetic substrates are correct. Therefore, more extended studies will be necessary to determine their clinical utility and significance under physiological and pathological conditions.

Use of negative staining technique and electron microscopy for the study of structural anomalies of outer dense fibres of human flagellum

Motility disorders due to tail defects are often seen in clinical andrology. Sperm motility should be assessed with regard to the morphology of the flagellum. Since suitable longitudinal sections are rarely obtained by routine transmission electron microscopy (TEM) and in view of the importance of dense fibres in modulating sperm motility and providing tensile strength, a detailed, study of human sperm flagellum by negative staining andTEM was attempted. The study was undertaken in two groups of men (I) fertile and (II) asthenozoospermic. The study revealed that outer dense fibres extend to 50–60% of the principal piece. Normal dense fibres were seen in 83% sperms and 23% sperms in groupsI andII respectively. The characteristics seen were variation in diameter, breakage or degradation with lacking or extended endpiece. The negative staining method provides an easy and useful analytical tool for identifying the defects of dense fibres and quantifying them.

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 (μm/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.

Clinical andrology.

Clinical andrology.

Recent advances in clinical/molecular andrology.

During the last decade there were extensive investigations in clinical and molecular andrology with emphasis on assisted reproduction, micromanipulation techniques of gametes, sperm/egg interaction, male contraception, diabetes mellitus, varicocele, andropause versus menopause, sexual dysfunction, associated hypertension/stress, prostatic carcinoma and molecular parameters of male reproduction. Sperm hyperactivation is a required step in capacitation sequence. Sperm motility is measured by videotape to evaluate the Straight Line Velocity (microm/s) (VSLI). Fertilization/embryonic development results from single sperm transfer (S-MIST) and multiple sperm transfer. Fertilization/embryo development is achieved by injection of immotile sperm into the perivitelline space. To assess sperm viability, a supravital stain suitable for use in combination with immunofluorescent assay, Hoeschst 33258, is used. The dye fluoresces with an intense blue when bound to DNA. To assess sperm plasma membrane integrity, a hypo-osmotic swelling test (HOST) is performed, using fluoresceinated D-mannose enriched albumin (FITC-DMA). The ability of sperm to swell under hypo-osmotic conditions indicates an intact membrane. A human protein, C-peptide, thought to be a useless byproduct of insulin may protect against devastating heart and nerve damage that diabetes causes. Human diabetics may benefit from the substance. Over 15 million Americans have diabetes, in which blood sugar levels rise out of control. There are two types of diabetics: Type I diabetics produce no insulin, the hormone that regulates blood sugar. Type II diabetics are unable to use their insulin properly. Diabetics are at great risk of heart disease and nerve damage, as arteries throughout the body leak and nerve-cell impulses fail. C-peptide is a byproduct of insulin production; it can be produced by the body or synthetically. Production of this protein is not induced by insulin, so diabetics who take insulin do not get C-peptide with it. Varicocele occurs unilaterally on the left side in 78% to 93% of men. Typically the presence of a varicocele is associated with an abnormal semen analysis (sperm density and morphology) and a decreased testicular volume on the affected side. Impaired sperm motility occurs in 89.5% of all varicocele patients. Varicocele ligation improves semen parameters in two thirds of patients. A few studies on andropause included sexual dysfunction, hormonal changes, medical/psychological correlates of impotence, ostenopenia/osteoporosis and bone loss; indices of bone remodeling, testosterone supplementation, androgen, negative feedback and hypothalamo-pituitary-testicular axis. Prostatic cancer is the second leading cause of cancer death for men between the ages of 60 and 80. Early detection involves a simple blood test for prostate specific antigen (PSA). Regular screening and early detection are essential. This is an important test because a high antigen count can be the only symptom. Since no screening is 100% accurate, physicians recommend both a PSA blood test and a physical examination. Although heredity plays a major role in whether a man will develop prostate cancer, men who lead healthy lives can dramatically reduce their chances of cancer: low-fat diet, eating plenty of fruits and vegetables and not smoking. Recent advances in molecular andrology include peptide hormone binding proteins; gonadotropin-releasing hormone (GnRH) agonists/antagonists analog; gonadotropins/their receptors; growth factors/reproduction; peptides as intratesticular regulators; molecular cloning of reproductive proteins/peptides. Gene cloning is applied for characterization/expression of genes coding. The interaction of gp120 with CD4 receptor plays a role in syncytium formation, apoptosis and CD4 cell deletion in human immunodeficiency virus (HIV) infection. The recombinant V3 peptide of fragment 307-330 of HIV-1 can induce sperm head agglutination. The generation process of react

Clinical andrology is important for treatment of male infertility with ICSI.

Clinical andrology is important for treatment of male infertility with ICSI.

Usefulness of the Acrobead Test in Evaluating Human Acrosome Function in Fresh and Cryopreserved Sperm

Purpose Assays for the acrosome reaction are usually cumbersome and lack reproducibility. Accurate determination of acrosomal status is important in patients diagnosed with male infertility before proceeding with intrauterine insemination or in vitro fertilization. We determined the optimum capacitation time and acrosomal status of fresh semen specimens in normal fertile men with the Acrobead † † Fertility Technologies, Inc., Natick, Massachusetts. test, and whether the assay could be used to evaluate cryopreserved semen specimens. Materials and Methods Semen samples from 13 normal donors were divided, with half of the fresh ejaculate used for the Acrobead test and half cryopreserved for a minimum of 24 hours in liquid nitrogen before testing. Fresh and frozen specimens were prepared with the swim-up technique. Sperm concentration was adjusted to 4, 2, 1 and 0.5 × 10 6/ml. in 4 wells of a 96-well tissue culture plate. Ten micro l. polyacrylamide beads (1.5 × 10 6/ml.) coated with anti-CD46 monoclonal antibodies (MH61 beads) were added to each well. The attachment of beads with acrosome reacted spermatozoa was scored after 0, 1, 3, 6 and 24 hours of incubation. Results were graded on a scale of 0 (no bead binding to the sperm) to 4 (complete attachment to the beads). Specimens with scores of at least 2 were considered normal. Results A score of at least 2 was noted in 3 of 13 fresh specimens (15.3%) at 1, 9 (69.2%) at 3, 11 (84.6%) at 6 and 13 (100%) after 24 hours. However, a significantly greater number of frozen specimens (8 of 13, or 62%) had a score of 2 or more at 1 hour of incubation and 100% bead attachment to sperm occurred at 3 hours. Conclusions Our results indicate that in fresh semen specimens an incubation period of 6 to 24 hours can be used to screen individuals who present with normal sperm characteristics but have slow acrosome reactions. Early acrosome reaction observed in cryopreserved specimens indicates that these spermatozoa may have membrane damage and leakage of acrosome contents as a result of the freeze-thaw process. The Acrobead assay is a simple and objective test that can be used at a clinical andrology laboratory to evaluate the acrosomal status of fresh but not frozen human spermatozoa.

Attitudes to clinical andrology: a time for change.

Attitudes to clinical andrology: a time for change.

HCG stimulation test for diagnosis of androgen deficiency.

A diagnostic-stimulation test with hCG (Pregnyl, Organon) was developed for demonstration of latent testicular deficit. The test was worked out on clinically healthy men and was used to diagnose cases of anorchidism and cryptorchidism. This stimulation test may be applied in clinical andrology as a noninvasive and highly sensitive method.

Treating male infertility needs more clinical andrology, not less.

Treating male infertility needs more clinical andrology, not less.

Laboratory standards in routine clinical andrology

It is a fundamental principle of laboratory tests that they are never entirely free from error. However, understanding the source and extent of such errors is a prerequisite for correct appreciation and interpretation of test results in the diagnostic process. In order to evaluate these errors, quality control (QC) has been introduced into clinical laboratory tests and has become routine practice.

Modern concepts of the structural bases for the reparative regeneration of the mammalian and human testis

Modern concepts of the structural bases of the reparative regeneration of the mammalian and human testis are discussed in this article, based on published data and our own findings. The significance of the rete testis as a regeneration zone is discussed as well as the use of its potentials in clinical andrology.

Assessment of human sperm acrosome reaction by flow cytometry: validation and evaluation of the method by fluorescence-activated cell sorting

To determine the applicability of flow cytometry to assess human sperm acrosome reaction. Prospective evaluation of semen samples incubated overnight for the development of spontaneous acrosome reaction or exposed to the calcium ionophore A23187 (5 microM) for 3 hours for induction of the acrosome reaction. University-affiliated tertiary care center. Normal healthy volunteers. The spermatozoa were stained with fluorescein isothiocyanate (FITC)-labeled pea agglutinin. The labeled samples were assessed visually and also subjected to analytic flow cytometry and fluorescence-activated cell sorting. Acrosome reaction assessed visually and by flow cytometry. Flow cytometric analysis showed a single peak of FITC fluorescence in the washed semen samples. A second peak of lower FITC fluorescence intensity was noted after overnight incubation or exposure to A23187, suggesting loss of fluorescence, which indicated the occurrence of the acrosome reaction. There was a statistically significant correlation between the assessment by the two methods (n = 41). However, although the mean difference between the methods was small (2.49%), the difference between the two methods for each individual sample can vary between -24% to +29%. When the sperm cells were subjected to cell sorting based on green fluorescence intensity, reanalysis and visual scoring verified that the low intensity peak contained a majority of acrosome-reacted spermatozoa (77.52% +/- 2.39%). These results validated the flow cytometric method for assessment of acrosome-reacted spermatozoa. Although flow cytometry is more objective and less time consuming when many samples are assessed at the same time, the visual method remains a useful and practical procedure in the clinical andrology laboratory.

Clinical andrology

Clinical andrology Get access Human Reproduction, Volume 7, Issue suppl_2, 1 January 1992, Pages 44–55, https://doi.org/10.1093/humrep/7.suppl_2.44 Published: 01 January 1992

Edward Martin (1859–1938). The founding father of modern clinical andrology

Edward Martin (1858-1938) was the first surgeon to treat successfully the problem of obstructive azoospermia, a feat that he first described in 1902. He subsequently carried out a total of 11 such operations that resulted in seven patencies and three pregnancies. He also demonstrated that the epididymis was the major site of these obstructive lesions. He pointed out the need for semen analysis in diagnosis and also demonstrated that azoospermia could have two causes, namely spermatogenic failure and ductal obstruction. He also made some important observations on sperm movement. He was probably the first clinician to demonstrate surgically ejaculatory duct obstruction, vasal atrophy and congenital absence of the vas. Most important of all, he demonstrated the need for accurate diagnosis in the treatment of male infertility. An argument is put forward that Edward Martin deserves the title of the 'Founding Father of Modern Clinical Andrology'.

臨床男性科学のめざすもの

臨床男性科学のめざすもの

BOOK REVIEWS

Complications of Gynecologic and Obstetric Management edited Michael Newton & Edward Newton. Multimodal Treatment of Ovarian Cancer Volume 20, Monograph Series of the European Organisation for Research and Treatment of Cancer edited Pier Franco Conte, Riccardo Rosso, Nicola Ragni & Jan B. Vermorken. Advances in Clinical Andrology edited C. L. R. Barratt & I. D. Cooke. Contraception: Science and Practice by Marcus Filschie & John Guillebaud. Fetal Monitoring. Physiology and Techniques of Antenatal and Intrapartum Assessment John A. D. Spencer. Baillière's Clinical Obstetrics and Gynaecology International Practice and Research Volume 3/Number 1 March 1989. Operative Treatment of Ovarian Cancer edited E. Burghardt J. M. Monaghan. Books received some of which may be reviewed at a later date Fetal and Neonatal Cardiology by Walter A. Long. Dictionary of Abbreviations in Medical Sciences by Rolf Heister. Perinatal and Multigeneration Carcinogenesis (IARC Scientific Publications No 96) edited by N. P. Napalkov, J. M. Rice, L. Tomatis & H. Yamasaki Ovarian Pathology (Current Topics in Pathology, Vol 78 edited by Franciso Nogales. A Guide to Effective Care in Pregnancy and Childbirth by Murray Enkin, Marc J. N. C. Keirse & Iain Chalmers. Viruses and Reproduction. A Bibliography compiled by Ernest L. Abel. Proceedings and Symposiums Fetal and Neonatal Physiological Measurements III (Proceedings of the Third International Conference on Fetal and Neonatal Physiological Measurements in Malmö/Ronneby, Sweden, 1988. Distributed by The Department of Obstetrics and Gynaecology, Malmo General Hospital, S‐21401 Malmo Sweden, 1989. Edited by Gerhard Gennser, Karel Marşscar;ál, Nils Svenningsen & Kjell Lindström

The sperm acrosome: functional and clinical aspects.

The acrosome of mammalian spermatozoa contains high amounts of acrosin, which is believed to be essential for gamete fusion, particularly for binding to, and penetration of, the zona pellucida. In addition, its activation from proacrosin seems to be associated with the capacitation process. Furthermore, it might facilitate cervical mucus penetration and intrauterine sperm migration by releasing kinins from kininogen, as well as participating in the acrosome reaction and in chromatin decondensation in the oocyte. Considering these functions of the acrosome during the process of fertilization, the morphology and functional integrity of the acrosome in ejaculated human spermatozoa are of fundamental importance in attachment, species-specific binding and zona penetration preceding gamete fusion. It is therefore mandatory to focus on acrosomal membrane functions and consider particularly the occurrence of acrosomal disturbances in spermatozoa from men of barren marriages. For this purpose, biochemical and immunocytochemical methods have been used to investigate acrosin activity and possible alterations of the acrosomal membrane system in different groups of sub- and infertile patients. Of particular interest is the ability of spermatozoa to undergo the acrosome reaction, which can be studied using the triple-stain technique. Of all the spermatological groups investigated (normo-, astheno-, oligo-, terato- and polyzoospermia), polyzoospermic semen samples showed severe acrosomal disturbances indicating functionally defective acrosomes which might hinder fertilization. The data indicate the need for the introduction of acrosomal markers in clinical andrology for proper diagnosis of acrosomal sperm defects.