Showing papers in "Andrologia in 2009"

Prevention of Fertility Disorders by Detection and Treatment of Varicocele at School and College Age

Abstract: Zusammenfassung Vorbeugung von Fertilitatsstorungen durch fruhzeitige Diagnostik und Behandlung der Varikocele bei Schulern und Studenten 4,067 Schuler und Studenten (12 - 15 Jahre) wurden auf das Vorhandensein einer linksseitigen Varikozele untersucht. Mit dem Einsetzen der Pubertat steigt die Haufigkeit von Varikozelen stufenweise an und bleibt danach konstant. In der gesamten Gruppe liegt die Frequenz bei 14.7%. Nur die Gruppen mit Varikozelen zweiten und dritten Grades konnen als “Risikogruppe” bezeichnet werden in Bezug auf eine spatere Unfruchtbarkeit; letztere wurde bei 5.3% aller Falle beobachtet. Allerdings entstehen schon bei einem Crittel der Jugendlichen mit einer Varikozele zweiten Grades und bei fast alien mit einer Varikozele dritten Grades anatomische Veranderungen (Volumen und Konsistenz) an den Hoden wahrend der Pubertat. Um einer eventuellen spateren Unfruchtbarkeit vorzubeugen, kann ein operativer Eingriff erwogen werden; jedoch mus man die psychologischen Reaktionen in diesem Alter mit besonderer Aufmerksamkeit berucksichtigen. Ein Eingriff im jugendlichen Alter zur Vorbeugung einer „moglichen” spateren Unfruchtbarkeit kann eine psychogene Impotenz auslosen. Resumen Prevencion de alteraciones de la fertilidad mediante detection y tratamiento del varicocele en la edad escolar Se examinaron 4.067 estudiantes entre 12 y 15 anos para ver si tenian varicocele izquierdo. Con el inicio de la pubertad la incidencia del varicocele aumenta gradualmente hasta que se completa la pubertad. Despues de la — adolescencia la incidencia de varicocele permanece constante. Para el grupo completo la incidencia de varicocele al-canzo el 14′7%. Puesto que solo los grupos con varicocele grados II o III, deben ser considerados como “grupos — riesgo”, en cuanto concierne a una infertilidad posterior, podemos establecer que el 5′3% de todos los estudiantes — pertenecen a este grupo. Por tanto un tercio de los sujetos con varicocele grado II y casi todos los sujetos con varicocele grado III, ya presentan en la adolescencia una influencia anatomica sobre el testicu-lo (volumen y/o consistencia). Puede considerarse la prevencion de una posible alteration posterior de la fertilidad, mediante tratamiento quirurgico del varicocele, pero es necesario una cuidadosa atencion por la reaction psicologica a esta — edad. La prevencion en la adolescencia de una posible alteration de la fertilidad en la edad adulta tiene el ries go de provocar reacciones psicologicas de inferioridad sexual y genital.

Standards for investigation of human semen Untersuchungsstandards für das menschliche Sperma La standardisation de l'analyse du sperme humain

Abstract: ummary A proposal for standardization of the analyses of human semen is presented with special emphasis on the necessity to perform accurate evaluation of sperm morphology, to determine the percentage of dead cells, and to carry out biochemical analyses of the seminal plasma to evaluate the secretory function of the accessory genital glands. Criteria for evaluation of sperm morphology as well as accuracy, precision and applicability of the various methods are presented. The proposed standards for evaluation of human semen constitute the basis for an already established collaboration between European andrologists aiming at a higher precision in semen analyses. Zusammenfassung Es wird eine Standardisierung der Spermaanalytik vorgeschlagen. Speziell wird eine genaue Ausarbeitung der Spermatozoenmorphologie gefordert, mit Angaben der Zahl toter Zellen. Dazu biochemische Analytik des Seminalplasmas um die sekretorische Funktion der akzessorischen Geschlechtsdrusen zu erfassen. Es werden genaue Methoden der Spermatozoenmorphologie beschrieben. Die angegebenen Standardisierungsvorschlage haben eine Zusammenarbeit vieler Europaischen Andrologen ermoglicht. Resume Une proposition de la standardisation de l'analyse du sperme humain est presentee en soulignant la necessite de realizer une determination exacte de la morphologie des spermatozoides, de mesurer le pourcentage de cellules mortes et de mener une analyse biochimique du plasma seminal pour evaluer l'activite secretoire des glandes genitales annexes. Les critees d'evaluation de la morphologie des spermatozoides aussi bien que la justesse, la precision et l'utilisation optimale de differentes methodes sent presentes. Les standards proposes de l'evaluation du sperme humain constituent la base d'une collaboration deja etablie entre andrologues europeens visant a une precision accure des analyses du sperme humain.

Spermadhesins: A new protein family. Facts, hypotheses and perspectives

Abstract: Spermadhesins are a novel family of secretory proteins expressed in the male genital tract of pig, horse and bull. They are major products of the seminal plasma and have been found to be peripherally associated to the sperm surface. The structure and function of spermadhesins have been thoroughly investigated in the pig, which exhibits the greatest diversity of members: AWN, AQN-1, AQN-2, PSP-I and PSP-II and its glycosylated isoforms. They are multifunctional proteins showing a range of ligand-binding abilities, c.g. carbohydrates, sulfated glycosamino-glycans, phospholipids and protease inhibitors, suggesting that they may be involved in different steps of fertilization. Isolated porcine spermadhesins bind the zona pellucida glycoproteins in a cation-dependent manner with a Kd in a low micromolar range, and AWN, AQN-1 and AQN-3 display similar binding affinity for glyoproteins containing Galβ(1-3)-GalNAc and Galβ(1-4)-GlcNAc sequences in O-linked and N-linked oligosaccharides, respectively. During sperm passage through the epididymis AQN-3 and AWN have been shown to bind tightly to the sperm surface by interaction with the phospholipids of the membrane bilayer. At ejaculation the spermadhesins form a protective coat around the sensitive acrosomal region of the sperm head, thus possibly preventing premature acrosome reaction. During in vitro capacitation most of these aggregated sperm adhesins are lost, with the exception of phospholipid-bound spermadhesins. AWN and AQN-3 may now serve as a primary receptor for the oocyte zona pellucida, thus contributing to initial binding and recognition between sperm and egg. The amino acid sequence of spermadhesins does not show any discernible similarity with known carbohydrate recognition domains (CRD). However, they belong to the superfamily of proteins with a CUB domain with a predicted all-β structure. The crystal structure of the heterodimeric complex of the spermadhesins PSP-I/PSP-II has been solved, showing that the overall structure of both spermadhesins consists of a β-sandwich with five (parallell and antiparallel) β-strands. It is the first three-dimensional structure of a zona pellucida-binding protein and reveals the architecture of the CUB domain. The spermadhesins represent a novel class of lectins that may be involved in sequential steps of fertilization, at least in the pig.

Subjective effects of Lepidium meyenii (Maca) extract on well-being and sexual performances in patients with mild erectile dysfunction: a randomised, double-blind clinical trial.

Abstract: Lepidium meyenii (Maca) is a cultivated root belonging to the brassica family used in the Andean region for its supposed aphrodisiac properties. We carried out a double-blind clinical trial on 50 Caucasian men affected by mild erectile dysfunction (ED), randomised to treatment with Maca dry extract, 2400 mg, or placebo. The treatment effect on ED and subjective well-being was tested administrating before and after 12 weeks the International Index of Erectile Function (IIEF-5) and the Satisfaction Profile (SAT-P). After 12 weeks of treatment, both Maca- and placebo-treated patients experienced a significant increase in IIEF-5 score (P < 0.05 for both). However, patients taking Maca experienced a more significant increase than those taking placebo (1.6 +/- 1.1 versus 0.5 +/- 0.6, P < 0.001). Both Maca- and placebo-treated subjects experienced a significant improvement in psychological performance-related SAT-P score, but the Maca group higher than that of placebo group (+9 +/- 6 versus +6 +/- 5, P < 0.05). However, only Maca-treated patients experienced a significant improvement in physical and social performance-related SAT-P score compared with the baseline (+7 +/- 6 and +7 +/- 6, both P < 0.05). In conclusion, our data support a small but significant effect of Maca supplementation on subjective perception of general and sexual well-being in adult patients with mild ED.

Testosterone-induced changes in testicular antioxidant system

Abstract: In order to investigate the role of testosterone propionate (TP) on the antioxidant system of the rat testis, lipid peroxidation (LPX) and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) of the testis of testosterone-treated and control rats were compared. The results indicate that TP administration to intact adult rats resulted in a significant decline in protein content of various subcellular fractions. This is accompanied with significant elevation in LPX levels of various subcellular fractions suggesting induction of oxidative stress. Activities of three enzymes related to the metabolism of superoxide radical (SOD) and hydrogen peroxide (CAT and GPx) of testis, were found to be significantly decreased in response to TP treatment. The role of testosterone in regulating testicular spermatogenesis through oxidative stress is discussed.

An Mg2+ and Ca2+-stimulated adenosine triphosphatase in human prostatic fluid--part II.

Abstract: Zusammenfassung Es wird uber die Enzym-Aktivitat eines Mg2+ und Ca2+-abhangigen ATPase-Enzym-systems berichtet, die an eine subzellulare Organelle geknupft und in der Prostata-Flus-sigkeit und damit im Seminalplasma vorhanden ist. Die elektronmikroskopische Beobachtung des uber 12 h bei 40 000 × G zentrifugierten Sediments aus der Prostata-Flussigkeit als auch aus dem Seminalplasma zeigte sekretorische Granulae und Vesiculae, wobei die meisten Partikel von einer dreischichtigen Membran umschlossen waren. Andere jedoch zeigten eine funfschichtige Membran. Das ATPase-Enzymsystem wurde charakterisiert; die scheinbare Michaelis-Konstante (Km) sowie Vmax der Mg2+-abhangigen ATPase ergab 7,1 × 10−4 M bzw. 4,0 mikromol pro 0,1 ml Prostata-Flussigkeit und 5 Min. Die entsprechenden Werte der Ca2+-abhangigen ATPase ergaben 3,8 × 10−4 M und 3,2 mikromol pro 0,1 ml und 5 Min. Concanavalin A fordert bei niedriger Konzentration das Mg2+-abhangige ATPase-System, indem es bei entsprechender Bedingung die Ca2+-abhangige ATPase-Aktivitat hemmt. Mehrere divalente Kationen (Cu2+, Zn2+, Mn2+ und Sr2+) sowie La3+ waren unterschiedlich hemmend. Cytochalasin B wirkte sich bei 5 × 10−5 M zu etwa 30% auf die ATPase hemmend aus. Vinblastin und Colchicin zeigten geringen oder keinen Einflus auf das System. Lipolytische Stoffe wie Deoxycholat, Triton-X-100 und Dodecanylsulfat waren auf das ATPase-System alle gleich stark wirkungsvoll als hemmende Substanzen. Auch Reaktoren der Thiol-Gruppen wirkten im gewissen Ausmas hemmend. Bei Zusatz von Na+ und K+ zum ATPase-System wurde kein Aktivitatsanstieg erhalten. Weiterhin ergab Ouabain keine Hemmung. Die partikulare ATPase-enthaltende Flussigkeit stammt sicherlich aus der Prostata. Verschiedene denkbare Funktiorien dieses ATPase-Systems werden diskutiert.

Effect of natural antioxidants, superoxide dismutase and hydrogen peroxide on capacitation of frozen-thawed bull spermatozoa.

Abstract: Bovine spermatozoa from frozen-thawed semen are sensitive to lipid peroxidation. Vitamin E protects sperm membrane against oxidative damage. Sperm capacitation produces structural changes on the plasma membrane. Reactive oxygen species could be involved in the capacitation process. The aim of this work was to study the influence of natural antioxidants on the plasma membrane and the influence of reactive oxygen species during bovine sperm capacitation. Sperm samples were frozen in a standard diluent, with and without vitamin E (1 mg ml-1). Heparin (60 micrograms ml-1) was used as a sperm capacitation inductor. Sperm capacitation was evaluated by chlorotetracycline assay. Lipid peroxidation was determined by the 2-thiobarbituric acid assay. A diminution of thiobarbituric acid reactive substances was observed in sperm samples frozen with vitamin E (P 0.05). When vitamin E and vitamin E + vitamin C were added to the capacitation medium, a significant decrease in the percentage of capacitated spermatozoa (P < 0.05) was observed in both cases. The addition of superoxide dismutase (0.1 mg ml-1) or H2O2 (50 microM) in the incubation medium, decreased the percentage of capacitated spermatozoa (P < 0.05). Vitamin E protects the plasma membrane against lipid peroxidation during sperm capacitation, and the presence of superoxide anion would be necessary for frozen-thawed bull sperm capacitation.

L‐ carnitine in idiopathic asthenozoospermia: a multicenter study

Abstract: The aim of the study described here was to evaluate any possible effect of L-carnitine on spermatozoal motility in a group of patients with unexplained asthenozoospermia in four different infertility centres. One hundred patients received 3 g d-1 of oral L-carnitine for 4 months. Sperm parameters were studied before, during and after this treatment. Motility was also studied by means of a computer-assisted sperm analysis. The results of the study indicate that L-carnitine is able to increase spermatozoal motility, both in a quantitative and in a qualitative manner (per cent motile spermatozoa increased from 26.9 +/- 1.1% to 37.7 +/- 1.1% [P < 0.001]; per cent spermatozoa with rapid linear progression increased from 10.8 +/- 0.6% to 18.0 +/- 0.9% [P < 0.001]; mean velocity increased from 28.4 +/- 0.6 microns s-1 to 32.5 +/- 0.8 microns s-1 [P < 0.001]; linearity index increased from 3.7 +/- 0.1 to 4.1 +/- 0.1 [P < 0.001], especially in the subgroup of patients with poor rapid linear progression of spermatozoa (per cent of motile spermatozoa increased from 19.3 +/- 1.9% to 40.9 +/- 1.4% [P < 0.001], and per cent of spermatozoa with rapid linear progression increased from 3.1 +/- 0.4% to 20.3 +/- 1.6% [P < 0.001]) An increase in spermatozoal output was also observed (total number of ejaculated spermatozoa increased from 142.4 +/- 10.3 10(6) to 163.3 +/- 11.0 x 10(6) [P < 0.001]). The authors conclude that oral administration of L-carnitine may improve sperm quality at least in patients with idiopathic asthenozoospermia.

Correlation between defects in chromatin condensation of human spermatozoa stained by aniline blue and semen characteristics.

Abstract: Human sperm heads which present disturbances of chromatin condensation are stained by acidic aniline, blue. To determine whether the proportion of unstained heads, i.e. with well condensed chromatin, can be considered as an index of sperm quality, a study was undertaken in 157 men during an infertility evaluation. In addition to the usual sperm characteristics, the percentages of unstained heads and of morphologically normal and abnormal forms were concomitantly evaluated. In a total of 15760 spermatozoa, the percentage of unstained heads was much higher in the population of morphologically normal forms than in that of abnormal forms (79.1% and 49.4% respectively, p less than 10(-9]. Among spermatozoa with structural abnormalities, it was much higher in cells with a single anomaly than in those with associated anomalies (53.9% and 40.6% respectively, p less than 10(-9]. When morphology was taken into account, only vitality was found to vary significantly with the percentage of unstained heads.

Morphology and functions of the human seminal vesicle

Abstract: The seminal vesicles originate in embryos of about 58 mm crown-rump-length from the Wolffian duct under the influence of testosterone. Along with the ampulla of the vas deferens and the ejaculatory duct, they form a functional unit that develops slowly until the onset of puberty. Developmental malformations occur as uni- or bilateral agenesis, aplasia, cysts, or ureterovesicular fistules. After puberty, the glands form sac-like structures which have a capacity of about 3.4-4.5 ccm and contribute about 70% of the seminal fluid. In addition to secretion, they are capable of reabsorption of fluids or dissolved substances, and of spermatophagy (ingestion and degradation of damaged spermatozoa by epithelial cells). Secretory activity of the glands is a measure of testosterone supplementation to the epithelium. Nervous regulation of secretion is realized by cholinergic post-ganglionic, sympathetic (and perhaps parasympathetic) fibres, derived from pelvic plexus. Contraction of the muscular wall occurs under the influence of excitatory adrenergic and modulatory NPY-encephalin-peptidergic nerve fibres. The secretory products of the seminal vesicles encompass (1) ions (K+: 1.1 mM ml-1) (2) low molecular weight substances (fructose: above 1.2 mg ml-1; prostaglandins above 250 microliters ml-1, (3) peptides (endorphin: 330 pg ml-1), and (4) proteins. In addition to plasma protein related forms such as transferrin, lactoferrin, and fibronectin, specific proteins such as semenogelin (52 kDa) are synthesized, the scaffold protein of semen coagulate forming the substrate for PSA (prostate specific antigen), sperm motility inhibitor (ca. 18 kDa), and others (placental protein 5, protein kinase inhibitor, carboanhydrase, 5'-nucleotidase), some of which are immunosuppressive. Therefore, functions of the seminal vesicles concern (a) formation of seminal coagulum, (b) modification of sperm functions (motility, capacitation), and (c) immunosuppression. Additional functions within the female genital system, perhaps during pre-implantation period, are likely, but remain to be proven experimentally.

über die Morphogenese rundköpfiger Spermatozoen des Menschen

Abstract: Zusammenfassung In einer andrologischen Praxis wurden bei einem infertilen Mann im Ejakulat ausschlieslich rundkopfige Spermatozoen beobachtet. Mit Hilfe der Semidunnschnitt-Technik und der Elektronenmikroskopie konnte an Ejakulat- und Biopsiematerial geklart werden, das in der Differenzierungsphase der Spermatogenese eine Misbildung am Akrosom auftritt: Das Akrosom entwickelt sich unabhaagig vom Kern im Zytoplasma der Spermatide, der Kern macht einen normalen Kondensationsprozes durch. Da das Akrosom somit keinen Einflus auf die Formgebung des Spermatozoenkernes ausubt, bleibt der Spermatozoenkopf rund. Im Ejakulat erscheinen nur akrosomlose Spermatozoen, die deshalb nicht befruchtungsfahig sind. Die cytologischen Besonderheiten bei der Differenzierung der Spermatide zum Spermatozoon werden im Einzelnen beschrieben und diskutiert. Resume Lors de l'examen d'un homme consultant pour sterilite les auteurs n'observent dans le sperme recueilli que des spermatozoides ayant une tete ronde. Le sperme et l'histologie testiculaire sont etudies en coupes semi-fines au microscope optique, en coupes ultra-minces au microscope electronique. Il a pu etre ainsi demontre qu'une malformation acrosomiale se developpe pendant la phase de differentiation de la spermatogenese: l'acrosome se developpe independament du noyau dans le cytoplasme des spermatides, le noyau luimeme subissant une condensation normale. Comme l'acrosome n'influence pas la morphogenese du noyau du spermatozoide il resulte que la tete du spermatozoide restera arrondie. Le sperme recueilli ne montre que des spermatozoides depourvus de leur acrosome et sont done non fertilisant. Les observations mises en evidence a propos de la differenciation des spermatides en spermatozoides font l'objet d'une description et d'une discussion detaillee.

Seminal reactive oxygen species-antioxidant relationship in fertile males with and without varicocele.

Abstract: The aim of this study was to assess seminal reactive oxygen species (ROS)-antioxidants relationship in fertile and infertile men with and without varicocele. One hundred and seventy six males were studied; fertile healthy volunteers (n = 45), fertile men with varicocele (n = 45), infertile oligoasthenozoospermia (OA, n = 44) without varicocele and infertile OA with varicocele (n = 42). In their seminal plasma, two ROS parameters (malondialdehyde, hydrogen peroxide) and five antioxidants (superoxide dismutase, catalase, glutathione peroxidase, vitaminE, vitaminC) were estimated. Compared with fertile healthy men, in all other studied groups, estimated seminal ROS were significantly higher and estimated antioxidants were significantly lower. Infertile men with varicocele showed the same relationship as infertile men without varicocele. Sperm concentration, total sperm motility as well as sperm normal forms were negatively correlated with seminal malondialdehyde and were positively correlated with vitaminC. It is concluded that varicocele has an oxidative stress (OS) in fertile normozoospermic bearing conditions. This may allow understanding that, within men with varicocele, there is a threshold value of OS over which male fertility may be impaired.

Polyunsaturated fatty acids in relation to sperm motility.

Abstract: The absolute amounts of phospholipid-linked fatty acids of different andrological conditions were analyzed by gas-liquid-chromatography. There was a significant linear correlation between docosahexaenoic acid and the spermatozoal density. Similarly there was a significant correlation between the content of docosahexaenoic acid and the number of motile normal sperm. These findings suggest that the process of lipid peroxidation is probably one of the biochemical causes of the low docosahexaenoic acid content in poorly motile sperm.

Effect of Cryoprotective Media and Dilution Methods on the Preservation of Human Spermatozoa

Abstract: Zusammenfassung Der Einflus von kryprotektiven Medien und Verdunnungsmedien auf die Erhaltung menschlicher Spermatozoen Menschliche Spermatozoen-Erhaltungs-Medium (HSPM), das aus 15% Glyzerin, 0,05 M Sucrose und 1% Glycin besteht, hat eine gleichwertige oder sogar bessere Kapazitat die Spermatozoen-Motilitat und - Vitalitat zu erhalten als Eigelbzitratmedium (ECM), welches 15% Glyzerin enthalt. Die Inseminationen mit dem Sperma, das in HSPM eingefroren wurde, hatten eine hohere Schwangerschaftsrate als wenn das Sperma in ECM eingefroren war. Glyzerin war DMSO oder Aethylenglykol als Kryoprotektants uberlegen, obwohl die kryoprotektive Fahigkeit von 7,5% Glyzerin und 7,5% Aethylenglykol nicht signifikant differierten. Weder die Zusatz-Temperatur noch die Methode des Zusatzes der Medien, die Glyzerin, DMSO oder Aethylenglykol enthielten, hatte irgendeinen Einflus auf die Sper-matozoen-Uberlebensart bei Tiefgefrierung. Raffinose (0,025 bis 0,05 M) oder Sucrose (0,05 bis 0,1 M) besserten die Uberlebensrate der eingefrorenen Spermatozoen. Der pH von HSPM hatte keinen signifikanten Einflus auf die Uberlebensrate der tiefgefrorenen Spermatozoen; der ideale pH lag zwischen 6,5 und 7,0. Das Verhaltnis von 1:1, 1:2, 1:3 und 1:4 des Verdunnungsmediums zum Sperma hatte keinen Einflus auf die Uberlebensrate der tiefgefrorenen Spermatozoen.

Influence of urogenital infections on sperm functions

Abstract: Many studies have examined the impact of genital tract infections on male fertility; however, the effect of bacteriospermia on sperm quality is still controversial. Bacterial infections are more frequently found in semen samples from asymptomatic infertile patients than in those from fertile men. Bacteriospermia is also a common problem of male partners from couples undergoing IVF. Therefore, the effects of microorganisms on human sperm acrosome reaction of oocytes have been studied in vitro and in vivo. Incubation of spermatozoa with Escherichia coli or Mycoplasma hominis in vitro resulted in reduced sperm motility and inducibility of acrosome reaction (delta AR) after exposure to calcium ionophore A23187. To show possible effects of E. coli and mycoplasma species on sperm functions in vivo, data from 488 patients were evaluated, in whose ejaculates microbiological examinations and determinations of acrosome reaction after exposure to low temperature had been performed. U. urealyticum and E. coli were found in semen samples from 52 and 31 men, respectively. M. hominis was only present in a minor number of samples and was not included in this study. Semen concentrations of E. coli and U. urealyticum ranged between 500-100,000 cfu x ml-1 and 100-80,000 cfu x ml-1. No correlation was found between delta AR and concentration of bacteria (Spearman rank correlation coefficient, E. coli: r-0.081, P = 0.6644; U. urealyticum: r = -0.081, P = 0.5698). In 69% of cases with U. urealyticum infection and reduced inducibility of acrosome reaction, this sperm function was normal after antibiotic therapy. However, improvement of acrosomal function may only be due to intra-individual variations of acrosome reaction. While E. coli and mycoplasma species affect sperm functions in vitro, the present data and a review of the literature fail to demonstrate similar effects in vivo.

Biochemical and Genetic Investigation of Round-headed Spermatozoa in Infertile Men including two Brothers and their Father

Abstract: Summary: Acrosin and the outer acrosomal membrane (OAM) were studied in the spermatozoa of 9 infertile patients who differed in the number of round-headed spermatozoa between 14 and 71% in their ejaculates. These sperm components were also investigated in two infertile brothers who exhibited exclusively round-headed spermatozoa in their ejaculates, and in their fertile father. It turned out that round-headed spermatozoa lack both acrosin and the OAM as studied by indirect immunofluorescent and immuno-peroxidase staining technique, gelatinolysis tests and by acrosin activity measurements. The normally shaped spermatozoa of 6 of the 9 infertile patients were found to be positive for acrosin and the OAM as expected, but in the remaining three patients even these spermatozoa were abnormal; in one patient they were unstainable for acrosin and in two patients they were unstainable both for acrosin and the OAM. These results have been confirmed by studies with the gelatinolysis test. The father of the two brothers with exclusively acrosomeless spermatozoa had more than 94% of normally shaped spermatozoa. However, only 10% of these spermatozoa were acrosin positive and only 30% were positive for the OAM. On the basis of these results we postulate that the mode of inheritance of the round-headed spermatozoa syndrome is polygenic rather than monogenic as suggested by previous authors. Zusammenfassung: Biochemische und genetische Untersuchungen bei Patienten mit Rundkopfspermatozoen Mit Hilfe spezifischer Antikorper gegen Akrosin und Proteine der aseren akrosomalen Membran wurden die Spermatozoen von 9 infertilen Mannern mit unterschiedlich vielen Rundkopfspermatozoen im Ejakulat untersucht. Diese Spermienkomponenten wurden auch analysiert bei 2 Brudern mit ausschlieslich Rundkopfspermatozoen im Ejakulat sowie bei deren Vater. Die Spermien der Patienten wurden ferner auf Akrosinaktivitat und gelatinolytische Aktivitat getestet. Es ergaben sich folgende Ergebnisse: Rundkopfspermatozoen reagieren nicht mit den Antikorpern gegen Akrosin und gegen die Proteine der auseren akrosomalen Membran, es fehlt ihnen die Akrosinaktivitat. Bei 3 der 9 Patienten und bei dem Vater des Bruderpaares reagieren auch die lichtmikroskopisch unauffallig aussehenden Spermien nur zum Teil mit den Antikorpern. Lediglich 10% bzw. 30% der Spermien des Vaters zeigen Immunfluoreszenz nach Behandlung mit dem Antikorper gegen Akrosin bzw. mit dem Antikorper gegen die ausere akrosomale Membran. Die Befunde legen den Schlus nahe, das das Syndrom der Rundkopfspermatozoen nicht monogen, sondern polygen vererbt wird.

Scavenging effect of N-acetyl-L-cysteine against reactive oxygen species in human semen: a possible therapeutic modality for male factor infertility?

Abstract: Summary. A new approach to reduce the level of reactive oxygen species (ROS) in human semen by using N-acetyl-L-cysteine (NAG) was evaluated. Semen samples were incubated with or without NAC (1.0 nig ml−1) at room temperature. The chemiluminescent signal of the oxidation of luminol was detected by means of an MTP reader after 0, 20, 40, 60 and 120 min, respectively, using 200 μM luminol. In addition, the dose-dependent action of NAC (0.1, 1.0 and 5.0 mg ml−1) and the influence of NAC on functional sperm parameters (motility and acrosome reaction) were studied. ROS levels decreased significantly after 20 min incubation with NAG. This reduction was greater in the high ROS group (>30 000 counts/107 viable sperm at t = 0) than in the low ROS group (<30 000). In addition, a marked dose-dependence of NAC was observed. Concerning sperm function, total sperm motility improved after incubation with NAC, but no significant change was observed with respect to the acrosome reaction. NAC (at concentrations of 1.0 mg ml−1) significantly reduced ROS in human semen and showed the possibility of improving impaired sperm function. After further testing NAC might be useful for the treatment of male infertility patients.

Rapid screening of the Y chromosome in idiopathic sterile men, diagnostic for deletions in AZF, a genetic Y factor expressed during spermatogenesis

Abstract: A rapid molecular screening programme has been established for the long arm of the human Y chromosome in Yq11 in order to quickly detect small interstitial deletions in this chromosome region. They have been observed in idiopathic sterile males with azoospermia and a severe oligozoospermia and are therefore indicative for deletion of AZF gene sequences. AZF (i.e. azoospermia factor) is a genetic factor located in Yq11 which controls human spermatogenesis. The screening programme is based mainly on a multiplex PCR approach using a series of Y-specific primers amplifying single DNA loci in Yq11. The order of all Y-DNA loci can be unequivocally arranged along the whole long Y arm. Therefore, any detected deletion can be quickly mapped in relation to the proposed position of AZF. Benefits and pitfalls of this new diagnostic Y screening method will be discussed.

Diabetic rat testes: morphological and functional alterations.

Abstract: Reproductive dysfunction is a consequence of diabetes, but the underlying mechanisms are poorly understood This study investigated the histological and molecular alterations in the testes of rats injected with streptozotocin at prepuperal (SPI rats) and adult age (SAI rats) to understand whether diabetes affects testicular tissue with different severity depending on the age in which this pathological condition starts The testes of diabetic animals showed frequent abnormal histology, and seminiferous epithelium cytoarchitecture appeared altered as well as the occludin distribution pattern The early occurrence of diabetes increased the percentage of animals with high number of damaged tubules The interstitial compartment of the testes was clearly hypertrophic in several portions of the organs both in SPI and SAI rats Interestingly, fully developed Leydig cells were present in all the treated animals although abnormally distributed Besides the above-described damages, we found a similar decrease in plasma testosterone levels both in SPI and SAI rats Oxidative stress (OS) is involved in the pathogenesis of various diabetic complications, and in our experimental models we found that manganese superoxide dismutase was reduced in diabetic animals We conclude that in STZ-induced diabetes, the altered spermatogenesis, more severe in SPI animals, is possibly due to the effect of OS on Leydig cell function which could cause the testosterone decrease responsible for the alterations found in the seminiferous epithelium of diabetic animals

Influence of different uropathogenic microorganisms on human sperm motility parameters in an in vitro experiment.

Abstract: Summary. The influence of different uropathogenic microorganisms (E. coli, enterococcus, Pseudomonas aeruginosa, Staphylococcus saprophyticus, Candida albicans) on human sperm motility was studied in vitro with a computer-assisted sperm analyser (CASA). Native ejaculates were prepared with the swim-up technique and adjusted to 22 times 106 spermatozoa ml−1. The sperm suspension was artificially infected with microorganisms in concentrations varying from 2 times 103 to 2 times 107. Sperm motility was examined directly after incubation, 2, 4 and 6 h later using the Mika motion analysis®, a computer-based, automatic motility analysis. Former results with E. coli (serotype 06) could be confirmed that a significant inhibitory effect on sperm motility was associated with bacterial growth. Experiments with the enterococcus strain and Staphylococcus saprophyticus indicated no significant influence on sperm motility parameters. Tests with Pseudomonas aeruginosa showed a decrease of progressive motility according to time, but not to different bacterial concentrations. A significant inhibitory effect of Candida albicans was only detected in the samples with the initial bacterial concentration of 2 times 107 microorganisms ml−1.

A New Method for the Treatment of Varicocele

Abstract: The development of techniques for selective catheterization of spermatic veins allowed unequivocal recognition of varicoceles including cases with minor or absent clinical symptoms and signs (Comhaire and Kunnen, Castro and Lima). The purpose of this communication is to demonstrate the feasibility of interrupting the retrograde flow in the spermatic veins by sclerosing the veins in the same intervention, without any significant prolongation of the diagnostic procedure.

Human Sperm Penetration into Zona-Free Hamster Oocytes as a Test to Evaluate the Sperm Fertilizing Ability

Abstract: Zusammenfassung Penetration menschlicher Spermatozoen in Zona pellucida-freie Hamster-Oozyten. Test zur uberprufung der Fertilisation von Spermatozoen Der Eintritt menschlicher Spermatozoen in pellucidafreie Oozyten von Hamstern wurde verwandt, um die Fertilitat menschlicher Samenproben einer andrologischen Klinik zu untersuchen. Die Chromatin-Dispersion des Spermas, im Phasenkontrastmikroskop festgestellt, wurde als Kriterium fur den Spermatozoeneintritt genommen. Die Ultrastrukturstudie zeigte, das das Verhalten der Gametenmembran wahrend der Verschmelzung sich nicht wesentlich von normalen Fertilitatsvorgangen unterscheidet. Vierundachtzig Proben wurden als normal (bezogen auf das Spermiogramm) eingeschatzt, aber nur 62 (74%) erzielten ein positives Ergebnis im Vergleich zu 30 (32%) positiven Ergebnissen von 95 Proben pathologischer Spermiogramme. Achthundertzehn Oozyten, die mit Spermatozoen von normalen Proben befruchtet wurden, erreichten eine Penetration von 25%, wahrend 1046 Oozyten, die mit pathologischen Samenproben befruchtet wurden, eine Penetration von 7% erlangten. Der Normalzustand der Hamsteroozyten wurde durch eine gleichzeitige Befruchtung mit Spermatozoen von Menschen und Hamstern nachgewiesen. Wahrend der Prozentsatz menschlicher Spermatozoenpenetration niedrig blieb, war die der Hamster uber 70%. Der vorliegende Bioassay wird als zusatzlicher Parameter zu den Spermiogrammen zur Untersuchung menschlicher Zeugungsfahgikeit empfohlen.

Association between sperm cell chromatin condensation, morphology based on strict criteria, and fertilization, cleavage and pregnancy rates in an IVF program.

Abstract: In this study, a total of 95 ejaculates from infertile patients were investigated morphologically according to Kruger's strict criteria and 78 of the 95 ejaculates were stained for chromatin condensation with acidic aniline blue. Patients were divided into two groups based on the percentage of morphologically normal spermatozoa as follows: Men with normal sperm morphology 14% (Group 2). The relationship between percentage of normal sperm morphology and fertilization, cleavage and pregnancy rate was analysed. The rates were 33.7%, 57.1% and 0.0% respectively, in the first group. The corresponding values for the second group were 76.1%, 68.2% and 32.1%. The fertilization and pregnancy rates correlate significantly with morphologically normal spermatozoa. In regard to the percentage of morphologically normal spermatozoa stained with aniline blue, patients were divided into two groups: patients with 0-20% stained spermatozoa (Group I) and those with > 20% (Group 2). Fertilization and pregnancy rates were higher in the first group than in the second group (79.9%, 52.8% vs. 58.8%, 29.5%). The results demonstrate that chromatin condensation visualized by aniline blue staining is a good predictor for IVF outcome and should be considered besides morphology by sperm assessment for patients undergoing IVF treatment.

Evoked sacral potentials in subjects with true premature ejaculation.

Abstract: Summary: Evoked sacral potentials were used to study the bulbo-cavernosus reflex in 85 patients with true premature ejaculation (TPE) and in 52 subjects as a control group. In the perineal and perianal recordings the amplitudes of the evoked responses (means ± SD) were 70 ± 91.1 μV and 35.6 ±36 μV in the TPE patients, and 39.2 ±36.8 μV and 26 ±21.6 μV in the controls, respectively. Both these differences resulted significant (P < 0.01 and P < 0.05, respectively). Our results suggest a reflex hyperexcitability, or an impaired “modulation” of the motor neurons of the pudendal nucleus by the regulating upper centers in the TPE patients. Evozierte sakrale Potentiale bei Patienten mit echter Ejakulatio praecox Zusammenfassung: Bei 85 Patienten mit einer echten Ejakulatio praecox wird der Bulbo-cavernosus Reflex mittels evozierter sacraler Potentiale gepruft. 52 weitere Manner dienten als Kontrollgruppe. Bei der perinealen und perianalen Aufzeichnung der evozier-ten Erregungen betrugen die Amplituden 70 ±91.1 μV und 35.6 ±36 μV bei den Patienten mit einer Ejakulatio praecox, bzw. 39.2 ±36.8 μV und 26 ±21.6 μV in der Kontrollgruppe. Bei beiden Gruppen sind diese Unterschiede signifikant. Die Ergebnisse weisen auf eine Reflexubererregbarkeit hin bzw. auf eine beeintrachtigte “Modulation” der mo-torischen Neuronen des Plexus pudendus durch die ubergeordneten Zentren bei Patienten mit einer echten Ejakulatio praecox.

Human testicular development and the role of the mesonephros in the origin of a dual Sertoli cell system.

Abstract: Zusammenfassung Die Hoden-Entwicklung des Menschen und die Rolle des Mesonephros bei der Genese eines zweiten Sertoli-Zell-Systems Einleitend wird eine kurze Ubersicht uber die verschiedenen Theorien zur Gonadenbzw. Hodenentwicklung gegeben: Die „klassische” Theorie, welche die somatischen Zellen vom Coelom-Epithel herleitet; die „mesenchymale” Theorie, die das Stutzzell- und das Zwischenzell-System ausschlieslich oder vorwiegend aus den Mesenchymzellen der Gonadenleiste entstehen last. Hinzu kommen die mehr funktionell orientierten Konzepte: Witschi's Theorie, nach der die Gonade sich aufgrund eines induktiven kortiko-medullaren Antagonismus zu Hoden oder Ovar differenziert und das Konzept von Jost, welches eine dyssymme-trische Entwicklung im Verlauf der sexuellen Differenzierung der Gonaden postuliert. Danach differenziert sich die Gonade aktiv unter genetisch bedingtem Einflus zum Hoden, wahrend sie ohne diesen Einflus zum Ovar wird. Gesteuert wird die Hodendiffe-renzierung durch einen Genort am Y-Chromosom, durch dessen Information ein H-Y Antigen (Histokompatibilitats-Antigen) produziert wird. Dieses Antigen determiniert die Hodenentwicklung. Ein weiteres chromosomales Regelsystem determiniert die hormonelle (androgene) Steuerung bei der Ausbildung der weiteren mannlichen Geschlechtsorgane und -merkmale. Im Hauptteil des Referates wird auf eigene Untersuchungsergebnisse uber die Rolle des Mesonephros (Urniere) bei der Genese der somatischen Zellelemente des Hodens (und Ovars) eingegangen. Bei der Differenzierung der Gonade zum Hoden (oder Ovar) werden somatische Zellen nicht nur vom Coelom-Epithel, sondern in betrachtlichem Umfang vom Mesonephros beigesteuert. Diese werden in einem intragonadalem Blastem (zentrales Blastem) gesammelt. Dieses Gonadenblastem ist beim Menschen im kranialen Pol der Hodenanlage (bzw. Ovar) lokalisiert, wo ein direkter Kontakt zu den degenerierenden Segmenten des oberen Mesonephrosabschnittes besteht. Zellen mesothelialer Herkunft (Coelom-Epithel) gelangen aus dem Bereich des oberen Coelomwinkels in das Blastem. Aufgabe des Blastems ist es, fur die fortgesetzte Bildung von Hodenstrangen auch nach der primaren Hodendifferenzierung Zellmaterial zu liefern. In den Strangen werden mit den primordialen Geschlechtszellen Stutzzellen (Pra-Sertoli-Zellen) eingeschlossen. Zwei Wochen nach der Hodendifferenzierung wird eine zweite Form von Pra-Sertoli-Zelle sichtbar, die vom Gonadenblastem uber die Strange des „Rete-Blastems” in die Hodenstrange gelangt und die Geschlechtszellen zur mitotischen Proliferation veranlast. Die Proliferation ist zeitlich koordiniert mit der praemeiotischen Proliferation der Oogonien im Ovar. Im Hoden leitet die Proliferation nicht in die Prophase der Meiose uber, da die hemmende Einwirkung der ersten Stutzzellpopu-lation zur Geltung kommt. Fordernde Zelle (meiosis-inducing Sertoli-cell) und hemmende Zelle (meiosis-preventing Sertoli-cell) konkurrieren in ihrem Einflus urn die Geschlechtszelle. Unmittelbarer Zellkontakt ist die Voraussetzung. Induzierende Zellen sind mesonephrogener, hemmende Zellen mesothelialer Herkunft. Eine ebenfalls mesonephrogene Herkunft wird fur die ubrigen somatischen Zellen des Hodens angenommen: Leydigzellen und peritubulare (myoide) Zellen.

Spermatophagy in the seminiferous tubules and excurrent ducts of the testis in Rhesus monkey and in man.

Abstract: Zusammenfassung Die Ultrastruktur von Zellen, die zur Phagozytose von Spermatozoen im Hoden und Nebenhoden befahigt sind, wurde mit Hilfe der Transmissions- und Rasterelektronen-mikroskopie untersucht. Es gibt ortsstandige Zellen, wie Sertolizellen und Retteepithelzellen und freie Makrophagen (Spermatophagen) in den Lumina der Samenkanalchen und Nebenhodenkanalchen, die Spermatozoen oder Bruchstucke von ihnen phagozytieren konnen. Die Spermatophagen entstehen aus Vorlauferzellen, die aus dem Hoden-Interstitium in die Tubuli seminiferi einwandern. Nach Phagozytose von Spermatozoen nehmen sie an Grose zu und enthalten manchmal mehrere Zellkerne. Grose abgerundete Spermatophagen sind im Nebenhoden zu finden. Beim Affen konnte festgestellt werden, das Spermatophagen im Bereich des Rete testis die Samenwege verlassen und in das subepitheliale Bindegewebe auswandern. Spermatophagie kommt gehauft vor bei alteren Mannern und in bestimmten Fallen von Oligozoospermie. Beim Affen treten zahlreiche Spermatophagen am Ende der spermatogenetischen Saison auf.

Improved staining method for differentiating immature germ cells from white blood cells in human seminal fluid.

Abstract: A new staining method for differentiating WBCs from immature germ cells in seminal fluid has been studied. It is a combination of Bryan's sperm stain, which particulary stains the acrosomal cap of the spermatozoa and the spermatid, and Leishman's blood stain which stains the WBCs in the same way as found in blood smears. The peroxidase positive granules in the cytoplasm of the PMN leukocytes are seen clearly. Thus, it is possible to differentiate PMN leukocytes from non-separated spermatids when they are present in a common cytoplasm. The staining of acrosomal cap permits differentiation between spermatids and lymphocytes.

Function of human epididymal proteins in sperm maturation

Abstract: Human post-testicular proteins were cloned by subtractive screening of epididymal cDNA libraries, employing testis as the primary negative control. This method identified six human epididymal cDNAs, named HE1-HE6, which are derived from abundant epididymal mRNAs. With the exception of HE5, which turned out to be identical to the lymphocyte surface antigen CD52, they represented completely novel human gene products. To date, there is little information on their function and the mechanism of their deposition on the sperm surface. Unlike the sperm coating antigens, CD52 binds firmly to the sperm membrane via its GPI anchor during epididymal passage. Its synthesis is carefully regulated by the epididymal epithelium. From the results of both in vivo and in vitro studies it was concluded that androgen and temperature are principal factors synergistically modulating epididymal CD52 expression. The human counterparts of two well-known major rodent epididymal proteins, secretory epididymal glutathione peroxidase (sGPX) and acidic epididymal glycoprotein (AEG = Protein DE), were not cloned by the subtractive screening approach, but by RT-PCR amplification.

Secular trend in reported sperm counts.

Abstract: Claims that there has been a recent secular decline in splerm counts have been disputed. Protaglonists on both sides of the discussion have based their arguments on data from a few large samples at different times and (mostly) in different places. It is not disputed that the differences between the mean counts in the various samples are statistically significant: what is a tissue is whether a secular decline is responsible. The present not adops a different approach to the problem: an attempt has been made to locate representative data on mean sperm counts of unselected men over the last 45 years. Twenty-nine such means are cited here. There can be no reasonable doubt that these reported mean sperm counts show a decline with time of publication, at least since 1960. It is not easy to see what artifact - other than a real secular trend - might responsible. Accordingly it is concluded that - at least in some places - it seems likely that a secular decline has occurred.

A double-blind trial of clomipramine in premature ejaculation.

Abstract: A double-blind clinical trial was designed to study the effect of clomipramine in 50 patients complaining of rapid ejaculation. Patients were divided into two groups and each group received 2 periods of 6 consecutive weeks' treatment with drug and placebo, so that comparison can be made between drug and placebo within each group and between the two groups. Analysis of collected data showed a significantly higher percentage of satisfactory performances with the drug than with placebo and maintenance of the beneficial effect after the drug was replaced by placebo.