bluuebunny/crossref_metadata_2025 huggingface dataset:
Gene expression in the dog epididymis: a model for human epididymal function
Authors: ELLERBROCK K. , PERA I. , HARTUNG S. , IVELL R.
Published: May 2008

The physiology of the epididymis is an integral part of the maturation process by which human spermatozoa acquire the abdity to reach and fertilize an oocyte. Because of the high degree of species specificity exhibited by the epididymal proteins involved in sperm maturation, we have assessed tissue from several alternative species for their suitability as a model for human epididymal physiology. Of these, the dog appears to offer an appropriate system. Northern hybridization using cDNA probes specific for human epidldymal genes established that, irrespective of dog breed, the canine equivalents of the epididymis‐specific HE1, HE4 and HE5 mRNAs were expressed highly in the canine epididymis. cDNA cloning and sequencing confirmed that the canine gene products, CE1, CE4 and CE5 were indeed true structural homologues of their human counterparts. Finally, tissue culture conditions were established wherein all three specific canine genes remained up‐regulated after 5 days of culture. Thus, the prerequisite criteria for the development of a system which models human epididymal physiology are to a large degree fulfilled by this canine culture system.
Quantified testicular histology in boys with sex chromosome abnormalities
Authors: MÜLLER JØRN , SKAKKEBÆK NIELS E. , RATCLIFFE SHIRLEY G.
Published: May 2008

Testicular histology in adult men with Klinefelter's syndrome (KS) is characterized by degenerative changes of the seminiferous epithelium. In contrast, limited changes have been reported in XXY fetuses. However, knowledge about the natural history of the degeneration of testicular histology in KS is scarce, and similarly testicular histology in prepubertal individuals with XY/XXY and XYY karyotypes is described incompletely. We have performed a qualitative and quantitative study of testicular histology in 11 XXY, one XY/XXY mosaic and two XYY boys between the neonatal period and 13 years of age using stereological methods and control data from normal XY boys and XY boys undergoing surgery for testicular maldescent. Testicular specimens were obtained at autopsy (n=2) or at time of surgery for malposition of the testis (n=10) or for inguinal hernia (n=2). The seminiferous tubules showed no signs of degeneration in any of the specimens. The volume density of seminiferous tubules was normal at all ages, whereas mean tubular diameter was reduced in all but three XXY boys. Germ cell numbers were only normal in XXY and XYY infants as well as in the 12 year old mosaic. No germ cells were observed in any XXY boy age 2 years or more. Leydig cells were observed in the infants and in a 13 year old XXY boy. The changes were comparable to what has been found in the most severely affected XY boys with testicular maldescent. We conclude that testicular histology in individuals with sex chromosome aneuploidy seems to be near normal during infancy, after which time degenerative changes occur.
A non‐human primate study (baboon; Papio hamadryas) to determine if a long‐acting progestogen, levonorgestrel butanoate, combined with a long‐acting androgen, testosterone buciclate, can suppress spermatogenesis: I. Dose‐finding study
Authors: GONCHAROV N. P. , KATZIA G. V. , BUTNEV V. U. , RICHKOR E. G. , MATIKAINEN T. , WAITES G. M. H.
Published: May 2008

This study in adult male baboons was conducted to establish the dose of a long‐acting progestogen, levonorgestrel butanoate, and that of a long‐acting androgen, testosterone buciclate, which, when combined, would achieve optimal and prolonged suppression of spermatogenesis. Two intramuscular injections of levonorgestrel butanoate at 3‐month intervals and in the dose range 1–8 mg/kg reduced sperm production and plasma concentrations of testosterone, LH and FSH for periods of up to 6 months. The suppression of sperm production was greatest and most prolonged in the 4 mg/kg group. Two intramuscular injections of testosterone buciclate at 3‐month intervals, and at doses of 4 and 8 mg/kg, induced variable changes in circulating levels of testosterone, elevated by the higher dose, and caused sperm suppression, in some animals to azoospermia. It was concluded that 8mg/kg testosterone buciclate would provide adequate androgen replacement when combined with 4 mg/kg levonorgestrel butanoate as a putative male contraceptive regimen.
A non‐human primate study (baboon; Papio hamadryas) to determine if a long‐acting progestogen, levonorgestrel butanoate, combined with a long‐acting androgen, testosterone buciclate, can suppress spermatogenesis: II. Efficacy study
Authors: GONCHAROV N. P. , KATZIA G. V. , BUTNEV V. U. , GORLUSHKIN V. M. , WAITES G. M. H.
Published: May 2008

Two combined injections of levonorgestrel butanoate (4 mg/kg) and testosterone buciclate (8 mg/kg) at 3‐month intervals to adult male baboons initiated a decrease in sperm concentration from baseline values of 490times106/ml to minimum values of 17times106/ml. This suppression was sustained until week 32, during which time between one and three azoospermic samples were collected from each of four out of five treated baboons in the period 10–24 weeks. Circulating plasma levels of LH and testosterone decreased to approximately 20–75% of baseline values. Plasma levels of cortisol declined to significantly reduced levels at weeks 22 and 36–44. All values returned to the baseline range by week 48. The combined administration of progestogen and androgen induced a more marked and sustained suppression to severe oligozoospermia or azoospermia than did the equivalent dose of progestogen alone (Goncharov et al., 1995). Although the dose of testosterone buciclate used did not maintain peripheral levels of testosterone in the normal range, it did not restimulate spermatogenesis. It was concluded that a combination drug regimen based on the novel long‐acting levonorgestrel and testosterone esters could provide a contraceptive option for men.
Bulk sperm collection by epididymal micropuncture and stimulation of the spermatic nerve: a novel method for sperm retrieval for IVF for surgically irreparable vasal obstruction
Authors: YAMAMOTO M. , HIBI H. , MIYAKE K. , FUKUGAKI H. , SUGANUMA N. , TOMODA Y.
Published: May 2008

We have developed a new sperm collection method by epididymal micropuncture in conjunction with perivascular nerve stimulation to obtain as many uncontaminated spermatozoa as possible for IVF for patients with surgically unreconstructable vasal obstruction. Fifteen azoospermic men with congenital absence of the vas deferens or vasal obstruction not amenable to surgical repair enrolled in this study. Under general anaesthesia the testis and epididymis were retrieved and stabilized in a testicular holder with the aid of 2% agar. The epididymal micropuncture was performed using a micropipette placed in a micromanipulator. A pair of Ag‐AgCl ring electrodes were placed around the spermatic cord for perivascular nerve stimulation with DC (intensity 136 V, frequency 20 Hz) for 30 s. Electrical stimulation was repeated every 1 min until no further fluid was obtained. Epididymal fluid obtained ranged from 78 to 980 μl (mean 169 μl). Sperm concentration ranged from 0.7 to 200 million (mean 53.5 million). Sperm motility ranged from 0.5% to 70% (mean: 19.9%). The percentage of spermatozoa with normal morphology ranged from 3% to 32% (mean 16.4%). Of 16 cycles in which spermatozoa and oocytes were retrieved, 56% (9/16) resulted in fertilization and 18.8% (3/16) developed clinical pregnancies. The clinical pregnancy rate per couple was 20% (3/15). This is the first clinical experience of the bulk sperm collection system for IVF for the treatment of unreconstructable obstructive azoospermia. Our results indicate that our new technique, in conjunction with IVF, is clinically useful for patients with surgically irreparable vasal obstruction.
Comparative analysis of proteins secreted in vitro by isolated seminiferous tubules from man and the rat
Authors: McKINNELL C. , BRACKENBURY E. T. , QURESHI S. J. , HARGREAVE T. B. , SHARPE R. M.
Published: May 2008

The aims of this study were to provide an overall comparison of the in‐vitro protein secretory profile of seminiferous tubules (ST) isolated from man and the rat, and to identify specific proteins secreted by both species. Two‐dimensional SDS PAGE was used to compare the profile of proteins secreted by cultured ST from 15 men undergoing orchidectomy with those secreted by ST isolated at stages VI‐VIII of the spermatogenic cycle from normal adult rats, and by ST at the same stages isolated from rats pretreated with methoxyacetic acid (MAA) in order to deplete both pachytene spermatocytes and round spermatids. Two abundant groups of proteins not present in the medium of rat ST were secreted consistently by human ST, though the profile of proteins secreted by human ST was otherwise more variable than in the rat. Twelve proteins secreted by ST isolated from humans were identified as possible homologues of rat ST‐secreted proteins, including the major rat Sertoli cell products SGP‐1 and SGP‐2, and an androgen‐regulated protein which derives in the rat from round spermatids. Otherwise, the majority of the 12 potential homologues corresponded to proteins which in the rat are secreted by ST from both normal and germ cell‐depleted testes or by ST from germ cell‐depleted testes only, suggesting that they are probably secreted by Sertoli cells and/or peritubular cells. Based on the potential homology of the proteins identified, our results suggest, first, that these proteins may play an important role in spermatogenesis and second, that the profile of proteins secreted by human ST is more akin to that secreted by ST isolated from germ cell‐depleted rats.
Phosphomannosyl receptors on the surface of spermatozoa from the cauda epididymis of the rat
Authors: BARBIERI A. M. , SOSA M. A. , GRIMALT P. , MAYORGA L. S. , BERTINI F.
Published: May 2008

This study demonstrates that β‐glucuronidase from rat preputial glands binds with high affinity to spermatozoa from the cauda epididymis. The binding was calcium‐independent and was inhibited by mannose‐6‐phosphate, but not by other phosphorylated or non‐phosphorylated sugars. Binding was also inhibited by α‐mannosidase from Dictyostelium discoideum, an enzyme known to have mannose‐6‐phosphate as the ligand. From solubilized sperm membranes, a protein of >200 kDa and one of 45 kDa, were adsorbed to a column of D. discoideum enzyme and to a phosphomannan column respectively, and eluted with mannose‐6‐phosphate. According to histochemical observations at the light and the electron microscopic level, gold particles coated with the enzyme became bound to the external surface of the plasmalemma in the acrosomal region of caudal spermatozoa. Similar labelling was observed using gold particles coated with antibodies against the rat 300 kDa phosphomannosyl receptor. The existence of phosphomannosyl receptors on the sperm plasma membrane, and our previous demonstration of the presence of affinity sites for epididymal β‐galactosidase on these gametes which is inhibited by phosphofructosyl derivatives, suggest strongly that maturing spermatozoa could be a target for glycosidases secreted into the lumen of the cauda epididymis, which then become bound to these cells via different ligand‐receptor systems.
Effect of testicular temperature on vasomotion and blood flow
Authors: SETCHELL B. P. , BERGH A. , WIDMARK A. , DAMBER J.‐E.
Published: May 2008

Vasomotion (spontaneous rhythmic variations in blood flow) has been demonstrated in the parenchyma of the testes of anaesthetized rats, using a laser‐Doppler flow probe. As the temperature of the testis was increased, mean blood flow showed no change, but the frequency of vasomotion increased and its amplitude decreased, until vasomotion disappeared between 36°C and 42°C. As the testis was then cooled, vasomotion reappeared, and increased in amplitude and decreased in frequency as the temperature fell. In control rats, in which the temperature of the testes was maintained at normal scrotal temperature of about 33°C, there were no changes in vasomotion over an equivalent period. In both groups of rats, when the temperature of the testes was then allowed to fall below normal scrotal temperature, the amplitude of vasomotion increased and its frequency decreased even further, without any change in mean blood flow. Capillary blood flow was also measured with microspheres at the end of the experiment, when testicular temperature was between 19°C and 27°C, and there was no difference between the cooled and control testes or epididymides.
Cholesterol, phospholipids and markers of the function of the accessory sex glands in the semen of men with hypercholesterolaemia
Authors: GRIZARD G. , SION B. , JOUANEL P. , BENOIT P. , BOUCHER D.
Published: May 2008

The effect of hypercholesterolaemia on the cholesterol and phospholipid content of spermatozoa and seminal plasma was studied. Testosterone and specific markers of the accessory sex glands were also measured. Semen samples from 11 hypercholesterolaemic patients (plasma cholesterol > 6.42 mmol/l, plasma triglycerides < 2 mmol/l) were compared with those of 11 normocholesterolaemic controls (plasma cholesterol < 5.14 mmol/l, plasma triglycerides < 2 mmol/l). Cholesterol, phospholipids and the molar ratio of cholesterol: phospholipids were not significantly different between the two groups of patients either in spermatozoa or in seminal plasma. In hypercholesterolaemic patients the total amount of carnitine in the ejaculate was significantly higher, but there were no significant differences in the levels of acid phosphatase or fructose. There were no significant differences in seminal plasma levels of testosterone in the two groups of subjects. These results suggest that hypercholesterolaemia has no effect on cholesterol and phospholipid levels in spermatozoa and does not cause gross modification of the secretory function of the accessory sex glands.
Effects of ischaemia on the caput epididymis and its relationship to higher epididymal obstruction: a qualitative study in the ram
Authors: MARKEY C. M. , JEQUIER A. M. , MEYER G. T.
Published: May 2008

The objective of this study was to determine if localized ischaemia of the caput epididymis in the ram causes morphological changes similar to those characteristic of higher epididymal obstruction in humans. This was tested by performing unilateral occlusion of the superior epididymal artery in 10 rams. At the end of 4 (n=5) or 28 weeks (n=4), rams were castrated and the testes and epididymides were weighed. Analysis of histological sections at the light microscope level provided quantitative data on lumen diameter and epithelial cell height of the efferent ducts. Qualitative analysis and specific histochemical stains for identification of lipofuscin pigment provided further information on tissue changes. Electron microscopy was performed on the efferent ducts to assess ultrastructural changes. The results revealed that localized ischaemia of the proximal epididymis caused a dramatic change in tubule calibre of the efferent ducts and initial segment resulting in obstruction of the distal lumina. These changes were more severe following 28 weeks of arterial occlusion. The epithelial cells of the proximal region showed an increase in the number of lysosomes and they became active in phagocytosis of spermatozoa. Lipofuscin pigment accumulated within the epithelial cells and also in macrophage‐like cells that had invaded the lumina, interstitium and intra–epithelial regions of the ducts. On the basis of these observations we conclude that the tissue changes which occur in the ram epididymis as a result of localized ischaemia show a striking similarity to those seen in men exhibiting higher epididymal obstruction. This suggests the possible implication of vascular disorders in the aetiology of obstructive azoospermia in men.
The origin of testicular microliths
Authors: NISTAL M. , MARTÍNEZ‐GARCÍA C. , PANIAGUA R.
Published: May 2008

The origin and development of eosinophilic bodies and microliths were studied histochemically and immunohistochemically in 29 human testes from prepubertal and adult males with different diseases. Non‐calcified and partially calcified eosinophilic bodies were found mainly in infants and children. Most of the eosinophilic bodies were apparently located within the seminiferous tubules, where they were surrounded by a double Sertoli cell layer. Serial sections revealed that these tubules were configurated abnormally, either displaying a spiral course or forming a cap‐like structure over the eosinophilic bodies that were actually extratubular. Completely calcified bodies (microliths) were found mainly in testes from adult men. The microliths were intratubular and appeared to be surrounded by a double layer of Sertoli cells. Eosinophilic bodies stained with eosin, periodic‐acid‐Schiff and silver methenamine like the tunica propria of seminiferous tubules. The peripheral band showed immunoreaction to both anti‐laminin and anti‐type IV collagen antibodies, similar to the basal lamina. Microliths were von Kossa‐positive and immunostained similarly to the eosinophilic bodies. The present results suggest that a microlith is formed as a small eosinophilic body in the tunica propria. Afterwards, the body increases in size by deposit of new substances and compresses the seminiferous tubule which forms a ring or cap around it. Thereafter, the body undergoes calcification and becomes a microlith which compresses the seminiferous epithelium and enters the tubule.
Investigation of some biochemical and functional effects of cryopreservation of human spermatozoa using an automated freezing–quick‐thawing method
Authors: MORSHEDI M. , OEHNINGER S. , BLACKMORE P. , BOCCA S. , CODDINGTON C. , HODGEN G.
Published: May 2008

The objective of the present studies was to assess the functional integrity of the sperm plasma membrane and metabolic and motility characteristics of the recovered motile fraction of human spermatozoa subjected to an automated freezing/quick‐thawing method. Sperm membrane features examined included progesterone‐induced changes in intracellular levels of calcium ([Ca2+]i), as measured by the fluorescent fura‐2 indicator, and the tight binding of spermatozoa to homologous zonae pellucidae as assessed by the hemizona assay (HZA). Basal [Ca2+]i, intracellular adenosine triphosphate (ATP) and adenosine diphosphate (ADP) levels, determined using chemiluminescence with luciferin‐luciferase, and motility parameters determined using a computer‐aided semen analyser (CASA) were studied concomitantly as an expression of metabolic/functional status. Ejaculates from fertile men (donors) were evaluated after swim‐up separation of the motile fraction in both fresh and cryopreserved–thawed samples, and fractions of each ejaculate (fresh and frozen–thawed) were subjected to parallel measurements of the same parameters at the same time frame. Basal and progesterone‐induced increase in [Ca2+]i, and ATP levels (up to 24 h) were similar in fresh and frozen–thawed samples. HZA results showed a modest (26%) although significant (p= 0.008) decrease in binding in frozen–thawed samples. The ratios of ATP/ADP in fresh and frozen–thawed samples were also found to be similar. Although post‐thaw sperm motility was significantly lower than that of the fresh samples, comparison of the results indicated that the method was capable of preserving > 65% of motile spermatozoa in almost all of the samples cryopreserved. Additionally, the swim‐up rescued a motile fraction in the frozen–thawed samples that was not significantly impaired with regard to motility, mean linear velocity or linearity as compared to the fresh fractions in the first 4 h. Our results show that this automated freezing–quick‐thawing method results in a small reduction in sperm–zona binding capacity, and that the time‐dependent decline in motility parameters observed for both fresh and cryopreserved–thawed samples cannot be related to ATP deficiency under the conditions of our experiments. These in‐vitro results are coincident with the maintenance of fertilizing capacity for donor spermatozoa in the in‐vitro fertilization (IVF) setting.
Stimulation of movement and acrosome reaction of human spermatozoa by PC12 liposomes encapsulating ATP
Authors: SKIBA‐LAHIANI M. , AUGER J. , TERRIBILE J. , FATTAL E. , DELATTRE J. , PUISIEUX F. , JOUANNET P.
Published: May 2008

The effect of co‐incubating human spermatozoa with 8 mmol/L dilauroylphosphatidylcholine (PC12) liposomes containing 6 mmol/L adenosine 5′‐triphosphate (LATP) was assessed by CASA and compared to that obtained with blank PC12 liposomes (LB). The aim of this study was to investigate if such treatments can improve sperm movement and sustain sperm motility over time. Significant and similar increases in straight‐line velocity and linearity of sperm movement in B2 capacitating medium (both p<0.01) were obtained with LB and LATP treatments (final concentration: 0.38 mmol/L PC12 and 0.5 mmol/L ATP) while in Tyrode's medium supplemented with 10 mg/mL BSA, these movement parameters were increased significantly only in sperm aliquots treated with LATP. Furthermore, after incubation for 0.5 h in Tyrode's, a bioluminescence assay of intracellular ATP indicated no significant change in ATP concentration for LATP‐treated spermatozoa while the ATP content of control and LB‐treated spermatozoa decreased significantly during the same period (both p<0.05). The effect of liposomes on the acrosome reaction was also investigated jointly with CASA. These experiments were performed by fluorescence microscopy, using PSA‐FITC and the supravital stain Hoechst 33258. After a precapacitation period of 3 h in BWW medium the spermatozoa were incubated for 1 h with LATP, LB, LB + free ATP and free ATP alone (final concentration 0.5 mmol/L ATP). Under these conditions the percentage of acrosome‐reacted spermatozoa was increased similarly after LATP and LB treatments compared to control (respectively from 4.9 to 12%, p<0.01 and 4.9 to 11.3%, p<0.05) but the percentage of true acrosome‐reacted spermatozoa, and the values for all movement characteristics (except percentage motility) were increased significantly only with LATP treatment. The results indicate the potential of PC12 vesicles for introducing highly hydrophilic compounds into spermatozoa, as well as for modulating membrane structures and functions required for fertilization.
Immunohistochemical quantitative study of the peritubular lamina propria after induction of testicular atrophy induced by epinephrine
Authors: SANTAMARÍA L. , MARTÍN R. , CODESAL J. , RAMÍREZ R. , PANIAGUA R.
Published: May 2008

Changes in the testicular peritubular lamina propria in rats treated for 1–11 weeks with intra‐scrotal injections of epinephrine were studied by quantitative immunohistochemical methods. In control testes, BrdU‐labelled nuclei (proliferating cells) were observed only in spermatogonia and some primary spermatocytes, whereas testes from epinephrine‐treated rats showed BrdU labelling in some of the spermatogonia and in peritubular cells. Immunostaining for transforming growth factor β (TGF‐β1) was present in germ cells, Sertoli cells and Leydig cells; vimentin immunostaining was found mainly in Sertoli cells; desmin immunostaining was found in the peritubular cells, and immunostaining for type IV collagen, laminin and fibronectin was found in the extracellular matrix of the lamina propria. The volume densities of seminiferous tubules (including seminiferous epithelium, lamina propria and tubular lumen) that immunostained for TGF‐β1, vimentin, laminin, desmin or fibronectin were calculated. All of these parameters increased significantly in testes from epinephrine‐treated animals during the course of the experiment, except for desmin immunostaining which showed no significant change in volume density. Since total seminiferous tubule volume decreased markedly in the testes of treated rats during the experiment, the transformation of relative values for immunostaining into absolute volumes per testis revealed a significant increase in TGF‐β1 immunostaining, no significant change in vimentin immunostaining, and a significant decrease in desmin immunostaining during the time of the study. The absolute volume occupied by laminin and fibronectin immunostaining decreased from the 3rd to the 8th weeks of treatment, and increased from the 8th to the 11th weeks. These changes, associated with germ cell depletion and tubular fibrosis, suggest that tubular ischaemic atrophy caused by epinephrine alters the peritubular myoid cells, which change immunophenotype and increase their secretion of the extracellular matrix components producing tubular fibrosis. The mechanism of this alteration may involve direct effects on the peritubular cells or the changes may be secondary to germ cell and/or Sertoli cell lesions.
European Academy of Andrology Prize, sponsored by Hamilton Thorne Research Ltd
Authors: MCKINNEY K. A. , BOYLE P. , THOMPSON W.
Published: May 2008

The objective of this study was to assess the in‐vitro effects of the nitric oxide substrate glyceryl trinitrate (GTN) on sperm motility and membrane lipid peroxidation. Nitric oxide (NO) can impair sperm motility, possibly by an alteration of cyclic nucleotide levels. NO may also be protective against lipid peroxidation. Semen samples from nine normospermic men were prepared by a swim‐up technique. Each specimen was divided into four aliquots, one of which was the control sample. The other three had 10‐6, 10‐8 or 10‐10 M GTN added. Sperm motility was then analysed over 180 min using a Hamilton Thorn motility analyser. Lipid peroxidation was assessed by measuring media malondialdehyde (MDA) levels at 180 min. Compared with control, the following measurements were reduced (p < 0.05) over the first 60 min in the 10‐6 M GTN aliquots only: mean path velocity (reduced by 14–15%), curvilinear velocity (reduced by 12–21%), straight‐line velocity (reduced by 18–19%) and percentage of hyperactivated spermatozoa (reduced by 38–43%). MDA levels and head movement parameters were comparable amongst all aliquots (p > 0.05). The depressant effects of GTN on sperm motility appeared to be transient and reversible. The effects observed may be due to NO generated by GTN, or to GTN itself. This suggests that NO may have a role in vivo as a physiological inhibitor of sperm motility. The addition of GTN did not appear either to cause sperm membrane damage or to protect the spermatozoa from lipid peroxidation.
Male germ cell transplantation in rats: apparent synchronization of spermatogenesis between host and donor seminiferous epithelia
Authors: JIANG F.‐X. , SHORT R. V.
Published: May 2008

Primordial germ cells (PGC) and gonocytes from male Sprague‐Dawlery rat fetuses and neonates were transplanted via the rete testis into the lumen of the seminiferous tubules of recipient adult Long Evans rats. The donor germ cells apparently differentiated into mini‐tubules or irregular segments of seminiferous epithelium within the lumen of the host seminiferous tubules, and exhibited qualitatively normal spermatogenesis in 10 out of 16 recipients. The stage of spermatogenesis of the intraluminal epithelium was synchronized closely with that of the adjacent seminiferous tubule epithelium, suggesting that the spermatogenic cycle is regulated locally by the intraluminal microenvironment. Male germ cell transplantation provides an interesting new tool for investigating the control of spermatogenesis.
Testicular size in young adult Sinhalese
Authors: WIKRAMANAYAKE E.
Published: May 2008

Height, body weight, arm span and testicular size were measured in 200 Sinhalese men aged 21–31 years. The mean ±SD right testicular size was 17.2 + 4.1 ml and that of the left was 16.1 + 3.9 ml. The testes were of equal size in 75% and the right testis was larger in 20% of the men. The prevalence of undescended testes was 2%. There was a significant correlation of testicular size with weight (p<0.05) and a significant correlation of right testicular size with span (p<0.01). Testicular size was not correlated with height or body mass index. Although the Sinhalese are ethnically closer to the Caucasians, their testicular size is similar to that reported previously for Thai men.
Ability of spermatozoa to bind to the zona pellucida during oligozoospermia induced with testosterone during a male contraceptive trial
Authors: LIU D. Y. , JOHNSTON R. , BAKER H. W. G.
Published: May 2008

To determine the ability of spermatozoa to bind to the zona pellucida (ZP) in testosterone‐induced oligozoospermia, previously fertile men participating in the World Health Organization (WHO) male contraceptive trial in Melbourne were studied while oligozoospermic to various degrees. Semen analysis was performed according to WHO methods. One or two ejaculates from each subject were cryopreserved before commencing weekly intramuscular injections of 200 mg testosterone enanthate. The frozen spermatozoa were used as controls for ZP‐binding tests of spermatozoa obtained during testosterone‐induced oligozoospermia (<10 times 106/ml) in either the suppression or efficacy (n=6) and recovery (n=3) phases. Two other subjects in the recovery phase with normozoospermia were also tested. Human oocytes that failed to fertilize in vitro from infertile patients were used for the sperm‐ZP binding test. Control (frozen) spermatozoa were labelled with fluorescein isothiocyanate and test (oligozoospermic semen) spermatozoa were labelled with tetramethylrhodamine B isothiocyanate. A mixture of equal numbers of labelled motile control and test spermatozoa were incubated with 4–6 ZP. There was a significantly (p <0.01) lower number of spermatozoa bound per ZP in oligozoospermic samples (65 ± 7, mean ± SEM) than in controls (80 ± 7). However, there were still large numbers of spermatozoa bound to the ZP for all the oligozoospermic samples. Five subjects had similar numbers of spermatozoa bound to the ZP for both control and oligozoospermic samples. Overall, the ZP‐binding ratio of test and control spermatozoa averaged 0.82 (range 0.51–1.13). Sperm‐ZP penetration was examined in three subjects and there were similar proportions of ZP penetrated for control (4/4, 4/4, 3/4) and oligozoospermic samples (4/4, 3/4, 3/4). In conclusion, although the number of spermatozoa bound to the ZP was significantly lower with testosterone‐induced oligozoospermia than with controls, the ZP‐binding ratio of oligozoospermic samples was within the normal range (>0.4). Also, the ability of spermatozoa to penetrate the ZP was not impaired. Thus, suppression of sperm production by testosterone appears to have little effect on the interaction between spermatozoa and the ZP.
Intrauterine insemination of washed spermatozoa for treatment of oligozoospermia
Authors: ARIBARG A. , SUKCHAROEN N.
Published: May 2008

Efficacy of intrauterine insemination (IUI) using washed spermatozoa for treatment of oligozoospermia was evaluated by a prospective randomized study in 50 couples, using LH‐timed natural intercourse in the alternate menstrual cycles as a control. The quality of spermatozoa in terms of their concentration and motility before and after sperm washing was compared. Sperm motility increased significantly after sperm preparation but the number of sperm was reduced. Eight pregnancies occurred in 253 cycles of IUI with washed spermatozoa and clomiphene citrate‐stimulated cycles (3.16% per cycle). Only one patient conceived in 242 LH‐timed natural intercourse cycles (0.41% per cycle). Compared with LH‐timed natural intercourse, IUI provided a significantly improved pregnancy rate. When the sperm count was <5times106 per ml, no pregnancy occurred with the IUI method. Therefore, IUI is of rather limited usefulness when the sperm quality is very poor. Few complications occurred after IUI, but included slight cervical contact bleeding and mild abdominal discomfort and/or cramps. In conclusion, IUI should be considered as a useful and relatively non‐invasive therapeutic modality for treating infertility caused by moderate oligozoospermia (>5times106/ml), when sexual intercourse fails.
Temporal changes in sulphated glycoprotein‐2 (clusterin) and ornithine decarboxylase mRNA levels in the rat testis after ethane‐dimethane sulphonate‐induced degeneration of Leydig cells
Authors: FRASOLDATI A. , ZOLI M. , ROMMERTS F. F. G. , BIAGINI G. , FUSTINI M. FAUSTINI , CARANI C. , AGNATI L. F. , MARRAMA P.
Published: May 2008

Short‐ (3–24 h) and long‐term (4–50 days) changes in sulphated glycoprotein‐2 (SGP‐2) and ornithine decarboxylase (ODC) mRNA levels in the adult rat testis were studied following a single dose of ethane‐dimethane sulphonate (EDS), to destroy the Leydig cells. Distribution patterns of SGP‐2 and ODC labelling were consistent with prevailing expression of the two transcripts in Sertoli cells and germ cells, respectively. This pattern did not show appreciable changes following EDS administration. No labelling of SGP‐2 mRNA was noted in the interstitium of control and EDS‐treated rats. This finding indicates that Leydig cell death induced by EDS is not associated with increased SGP‐2 mRNA levels, a phenomenon related to apoptotic cell death in many tissues. Semi‐quantitative densitometric analysis of the preparations demonstrated differential changes in SGP‐2 and ODC mRNA levels in the tubular compartment following EDS treatment. At 6, but not at 3 and 12, h following EDS administration, SGP‐2 mRNA levels showed a significant increase, possibly secondary to a direct effect of the alkylating agent on Sertoli cells. A significant decrease in ODC mRNA levels was observed from day 7 to day 28, matching degenerative changes in the seminiferous epithelium. In contrast, a decrease in SGP‐2 transcript levels was observed from days 21–35 after treatment.

In conclusion, our findings demonstrate that SGP‐2 mRNA, a putative marker of apoptosis, is not altered in the testicular interstitium during EDS‐induced degeneration of Leydig cells. In the tubular compartment, the content of both ODC and SGP‐2 mRNAs following EDS administration appears to be dependent mostly on the integrity of the germ cell complement and not to be influenced directly by changes in testosterone levels.
Validation of a scoring method predicting the in‐vitro fertilizing ability of human spermatozoa
Authors: PARINAUD J. , RICHOILLEY G. , MILHET P. , MOUTAFFIAN H. , VIEITEZ G.
Published: May 2008

The fertilizing ability of human spermatozoa depends upon numerous functions such as motility, normal morphology, ability to bind to the zona pellucida and to undergo the acrosome reaction. Hence a lot of tests have been developed to try and predict IVF results. In a previous study we had established a scoring method, based on parameters such as sperm morphology, vitality, motility and the acrosome reaction, which was able to predict up to 83% of in‐vitro fertilization results. The present study aimed to validate this score on a separate set of sperm samples. The results confirmed those of the first series. The score allowed prediction of fertilization failures with a 56% sensitivity, a 91% specificity, a 56% positive predictive value and a 91% negative predictive value. Therefore, this score could be used routinely to choose between conventional IVF and ICSI.
Electrovasography in normal and vasectomized men before and after vasectomy reversal
Authors: SHAFIK A.
Published: May 2008

The electrical pattern of the vas deferens, or electrovasogram (EVG), was studied in 22 healthy volunteers, 20 vasectomy subjects and 18 individuals after vasectomy reversal. Their mean ages were 38.8 ± 7.3, 44.3 ± 7.9 and 58.6 ± 6.6 years, respectively. Two electrodes were applied to the posterior aspect of the upper part of the scrotum. During the operation of vasectomy reversal, an electrode was applied directly to each of the two vasal segments before and immediately after vasovasostomy. The electrical activity and intravasal pressure were measured. In normal subjects, slow waves or pacesetter potentials (PP) were recorded. They had identical frequency and a regular rhythm from the two electrodes and were consistent in the individual subject on all test days. PP were followed randomly by bursts of action potentials (AP). In vasectomized subjects, PP from the proximal vasal segment exhibited an irregular rhythm (vasoarrhythmia). During operation for vasectomy reversal, the proximal vasal segment recorded vasoarrhythmia whereas the distal segment revealed a silent EVG. Intravasal pressure was normal (p > 0.05) in the distal segment but high (p < 0.05) in the proximal segment. EVG performed 1–6 years after vasectomy reversal showed a normal pattern in four subjects and diminished frequency and amplitude in three. These seven subjects had impregnated their wives and had an obstructive interval of < 3 years. The 11 subjects who did not produce a pregnancy had a vasoarrhythmic EVG and an obstructive interval of > 3 years. In conclusion, an EVG could be identified for normal subjects. Vasectomy resulted in a vasoarrhythmic EVG pattern which proved to be correctable by vasectomy reversal if the obstructive interval was short.
The combination of two semen preparation techniques (glass wool filtration and swim‐up) and their effect on the morphology of recovered spermatozoa and outcome of IVF‐ET
Authors: AL‐HASANI S. , ALPÜSTÜN S. , LUDWIG M. , DIEDRICH K. , BAUER O. , KÜPKER W. , WOLFF A. , KREBS D.
Published: May 2008

The aim of this prospective study was to relate sperm quality, especially sperm morphology, to the outcome of IVF results, when glass wool filtration (GWF) and swim‐up were used in combination for the preparation of spermatozoa. A total of 60 ejaculates were analysed. GWF was compared with glass wool filtration/swim‐up (GWF‐SU) by using aliquots from the same semen samples to increase the precision of the comparison and to establish the cumulative effect of these two semen preparation methods on the morphology of the recovered spermatozoa. Sperm parameters were examined in native semen, in semen preparation samples after GWF and GWF‐SU. The mean percentages for motility, morphology and velocity were improved significantly over those in fresh semen only when GWF‐SU was used. GWF alone resulted only in a significantly higher recovery of motile spermatozoa. In semen preparation, 10% of spermatozoa with normal morphology appeared to be the cut‐off point as there was a significantly higher recovery in fresh semen samples containing > 10% morphologically normal spermatozoa after GWF‐SU. Outcome of IVF‐ET following preparation with GWF‐SU showed better results in comparison to the swim‐up procedure alone, though this difference was not statistically significant. The cut‐off point was also > 10% morphologically normal spermatozoa. Improvement in all aspects of IVF‐ET occurred when native semen contained > 10% morphologically normal spermatozoa. It is concluded that GWF alone did not produce significantly better results but, in combination with swim‐up, it resulted in significantly better sperm morphology and in an improved outcome of IVF‐ET for fresh semen samples with > 10% morphologically normal spermatozoa.
RU486 inhibits penetration of human spermatozoa into zona‐free hamster oocytes
Authors: YANG J. , SERRES C. , BAULIEU E.‐E. , JOUANNET P.
Published: May 2008

Previous studies have shown that RU486 decreases the concentration of intracellular calcium ([Ca2+]i) in human spermatozoa in vitro and partially antagonizes the effect of progesterone on calcium influx and sperm acrosome reaction. The present study has examined the effect of RU486 on the penetration of human spermatozoa into zona‐free hamster oocytes. RU486 (10 μM) decreased significantly the rate of penetration of zona‐free hamster oocytes by human spermatozoa, and nearly abolished penetration when present in the medium at concentrations of 50 and 100 μM. RU486 must be present in the medium to exert its inhibitory effect. At the same concentration (10 μM) as RU486, progesterone was unable to reverse the inhibitory effect of RU486 on the penetration rate of human spermatozoa.
Glucose is not essential for the occurrence of sperm binding and zona pellucida‐induced acrosome reaction in the mouse
Authors: URNER F. , SAKKAS D.
Published: May 2008

Glucose is necessary, in some species, for successful fertilization, but its role in the different gamete functions has only been documented partially. In this study we have examined the progression of mouse spermatozoa through the oocyte investments in order to determine which steps require glucose. Evidence is presented of glucose involvement during this sequence of events, since in‐vitro fertilization was inhibited in glucose‐free medium even when spermatozoa were capacitated in the presence of glucose. Access to the zona pellucida was not inhibited in the absence of glucose, suggesting that capacitation, progressive motility and zona binding of spermatozoa do not depend on the presence of an extracellular glycolysable substrate. However, zona penetration was prevented in the absence of glucose. Most importantly, the zona pellucida‐induced acrosome reaction was found to occur in glucose‐free medium. It is proposed that hyperactivated motility, which has been described to require glucose in the literature, may be the precise process responsible for the failure of spermatozoa to enter the perivitelline space.
Reproductive parameters of older compared to younger men of infertile couples
Authors: ROLF C. , BEHRE H. M. , NIESCHLAG E.
Published: May 2008

In order to evaluate possible age‐dependent influences on male infertility, semen parameters, sex hormone levels and pregnancy rates were compared retrospectively in three groups of men attending our fertility clinic: group A: 39 patients > 50 years. Group B (39 patients < 30 years) was established as the ‘standard infertile patient’ seen in our Institute. These patients were matched with patients of group A according to the year of their first visit to the clinic. Group C: 39 young patients (< 30 years) who were selected on the basis of their wives' advanced age. The frequency of sexual intercourse in group A was significantly lower compared to group C. The period of sexual abstinence was longer in group A than in group B. Ejaculate volume and sperm motility in group A were lower compared to groups B and C. The differences in other semen parameters were only minor. The differences in serum levels of testosterone and FSH were significant between group A and groups B and C. Pregnancies were reported in 6 of 26 couples from group A (duration of infertility: mean 79 months, range 16–187), in 17 of 28 couples from group B (duration of infertility: mean 54 months, range 16–104) and in nine of 30 couples from group C (duration of infertility: mean 66 months, range 25–125). Since the infertility pattern of me older patients desiring offspring was not significandy different from that of younger patients, it is concluded that the age of the female partner, rather than the age of the husband, is most important in determining the fertility chances of couples presenting with infertility.
A placebo‐controlled flow cytometric study of the effect of low‐dose prednisolone treatment on sperm‐bound antibody levels
Authors: RÄSÄNEN M. , LÄHTEENMÄKI A. , AGRAWAL Y. P. , SAARIKOSKI S. , HOVATTA O.
Published: May 2008

In this placebo‐controlled study the effect of short term, low‐dose prednisolone treatment on sperm‐bound antibody levels was investigated using an objective flow cytometric (FCM) measurement. Twenty infertile men, positive for antisperm antibodies by the mixed antiglobulin reaction (MAR) test, were treated with either 20 mg/day prednisolone (n= 10) or placebo (n= 10) for 3 weeks. The two groups were similar with respect to sperm count, motility and antibody levels. The proportion of spermatozoa positive for IgG and IgA antibodies and the antibody load, expressed as the number of IgG and IgA antibody molecules bound per spermatozoa were measured by FCM. The study showed that, overall, the antibody levels measured before and after treatment with low‐dose prednisolone or with placebo were not statistically different. However, in the prednisolone group a marked decrease in the proportion of spermatozoa positive for IgG and IgA antibodies was observed in two of the patients. An examination of the FCM data of these two patients revealed that, in their ejaculates, only the spermatozoa with low to moderate amounts of sperm‐bound antibodies had responded to prednisolone treatment, while spermatozoa with high levels of sperm antibodies had remained resistant to prednisolone. These results point to the existence of an as‐yet‐undefined subset of prednisolone‐responsive, antibody‐positive spermatozoa.
Glutathione S‐transferase‐M1 ‘null’ genotype and alcohol‐induced disorders of human spermatogenesis
Authors: PAJARINEN J. , SAVOLAINEN V. , PEROLA M. , PENTTILÄ A. , KARHUNEN P. J.
Published: May 2008

The association between alcohol‐induced disorders of human spermatogenesis and glutathione S‐transferase‐M1 genotype was investigated in an autopsy study comprising 271 subjects, including interviews with relatives or close acquaintances on the alcohol consumption of the deceased. Of the 50 moderate drinking men (reported mean daily alcohol consumption < 40 g), 21 (42.0%) had normal spermatogenesis, whereas 24 men (48.0%) had partial, and five (10.0%) complete arrest of spermatogenesis. Of the 21 men with normal spermatogenesis, nine (42.9%) had GST M1 ‘null’ genotype, while this genotype was found in 13 (44.8%) of the 29 men with partial or complete spermatogenic arrest. Of 212 heavy‐drinking men (reported mean daily alcohol consumption > 80 g), only 45 (21.2%) had normal spermatogenesis, whereas 77 (36.3%) had partial spermatogenic arrest. Complete arrest of spermatogenesis was found in 81 men (38.2%) and nine men had Sertoli‐cell‐only syndrome (4.2%). Of the 45 heavy drinkers with normal spermatogenesis, 27 (60%) men had GST M1 ‘null’ genotype (OR 2.7, with 95% confidence intervals: 1.0–4.0, when compared to those with disorders of spermatogenesis). The frequency of GST M1 ‘null’ genotype in heavy drinkers with normal spermatogenesis also differed from that of corresponding moderate drinkers, whereas the frequency of GST M1 ‘null’ genotype in heavy drinkers with disorders of spermatogenesis was similar to moderate drinkers with or without disorders of spermatogenesis. The finding that > 20% of men in the heavy‐drinking group had normal spermatogenesis suggests individual variations in sensitivity to alcohol‐induced disorders of this process. Heavy drinkers with GST M1 ‘null’ genotype were slightly less prone to develop disorders of spermatogenesis. Thus, the GST M1 locus may be associated with susceptibility to develop alcohol‐induced disorders of spermatogenesis.