bluuebunny/crossref_metadata_2025 huggingface dataset:
Testicular blood flow and vasomotion can be maintained by testosterone in Leydig cell‐depleted rats
Authors: DAMBER J.‐E. , MADDOCKS S. , WIDMARK A. , BERGH A.
Published: May 2008

The effect of testosterone supplementation on testicular blood flow, testicular vasomotion, the number of polymorphonuclear leucocytes (PMN's) in testicular blood vessels and prostatic blood flow were studied in rats in which the Leydig cell had been destroyed specifically by a single injection of ethane dimethylsulfonate (EDS). Other rats were supplemented with testosterone by subcutaneous injection of 25 mg testosterone propionate on days 1, 3 and 6. In some experiments, the effect of a single injection of 25 or 125 mg testosterone was studied. Testicular and prostatic blood flow and the number of PMN's in testicular blood vessels decreased, and vasomotion disappeared in Leydig cell‐depleted rats, but testosterone supplementation restored all parameters to normal values. Moreover, a single injection of testosterone was able to restore testicular and prostatic blood flow to normal levels but had an inconsistent effect on vasomotion. These results suggest that testosterone may play a role in the physiological control of the testicular microcirculation.
Purification and characterization of phospholipase A2 from bovine prostate
Authors: RÖNKKÖ S.
Published: May 2008

Phospholipase A2 (PLA2) was purified from bovine prostate by ammonium sulphate precipitation and fractionation by anion exchange chromatography, chroma‐tofocusing and gel filtration. The purified enzyme was Ca2+‐dependent and had a pH‐optimum of 8.0. Ba2+, Fe2+, Hg2+, Mg2+, Pb2+, Sr2+ and Zn2+ as well as lysophosphatidylcholine, lysophosphatidylethanolamine and p‐bromophenacyl bromide (p‐BPB) inhibited the enzyme strongly. The enzyme had an estimated molecular weight of 12000 ± 1000 daltons on SDS‐PAGE. Isoelectric focusing showed one PLA2 activity‐containing band at pl 5.3. The purified enzyme hydrolysed linoleic acid at the sn‐2 position of phosphatidylcholine and phosphatidylethanolamine with high selectivity, compared to arachidonic acid.
Immunohistochemical demonstration of androgen receptors on testicular blood vessels
Authors: BERGH A. , AMBER J.‐E. D.
Published: May 2008

Using an antiserum directed against the human and rat androgen receptor we have used immunohistochemistry to demonstrate that nuclear androgen receptors are present in the muscular layer of almost all arteries within the rat testis. These receptors are apparently influenced by testosterone. They disappear after Leydig cell depletion induced by ethanedimethane sulphonate (EDS) and return after testosterone treatment. It is suggested that testicular blood vessels could be a target‐organ for androgens and may mediate some of the effects of androgens on the testicular microcirculation.
Castration‐resistant secretion in the hamster seminal vesicle does not depend on androgens
Authors: PINHO M. S. , MATA L. R.
Published: May 2008

The secretory activity of seminal vesicles (SV) in the castrated hamster was studied by stereological analysis and biochemical approaches following treatment with cyproterone acetate (CPA) and adrenalectomy in order to investigate whether extra‐testicular androgens are responsible for castration‐resistant protein secretion.

Treatment of castrated animals with CPA decreased the size of secretory granules and increased the number of apical granules, though neither the absolute nor the relative volume of all the components analysed was changed. In addition, CPA‐treatment increased the amount of protein exocytosed by SV in castrated animals, though total protein synthesis remained unchanged. Adrenalectomy neither suppressed secretion nor induced any further ultrastructural changes in the SV epithelium.

Our results demonstrate that secretory activity of the hamster SV following castration is not controlled by extra‐testicular androgens and suggest that SV secretory proteins, which are heterogeneous with regard to their sensitivity to androgen withdrawal, might be regulated diferentially by androgens.
A positive role for the superoxide anion in triggering hyperactivation and capacitation of human spermatozoa
Authors: Lamirande E. De , Gagnon C.
Published: May 2008

Capacitation of human spermatozoa is essential for fertilization, and is characterized visually by hyperactivated motility. We have investigated whether reactive oxygen species could induce these two events in human spermatozoa. The addition of xanthine + xanthine oxidase + catalase (X+XO+CAT: generation of superoxide anion and removal of hydrogen peroxide) and foetal cord serum (FCS), a known biological inducer of capacitation and hyperactivation, to spermatozoa, induced levels of hyperactivation (15.4 ± 1.6% and 8.0 ± 1.0%, respectively) which were significantly higher than that of controls (5.4 ± 0.6%). The hyperactivation measured was part of the capacitation process. Furthermore, the addition of superoxide dismutase prevented the capacitation and hyperactivation induced by X+XO+CAT or by FCS. These results suggest that the superoxide anion may be involved in capacitation and hyperactivation of human spermatozoa.
Enhancement of testosterone secretion by normal adult human Leydig cells by co‐culture with enriched preparations of normal adult human Sertoli cells
Authors: Lejeune Herve , Skalli Mourad , Sanchez Pascale , Avallet Odile , Saez Jose Maria
Published: May 2008

An in‐vitro method was developed to study Sertoli—Leydig cell interactions in man, using testes removed at the time kidneys were removed for transplantation from 6 young adult men (aged 17–45 years) after cerebral death. After collagen‐ase digestion of testicular tissue, Leydig cells were purified on discontinuous Percoll gradients. Two fractions of Leydig cells, ‘L2’ and ‘L3’ which differed in their buoyant density (1.05 g < L2 < 1.06 g and 1.06 g < L3 < 1.08 g), were obtained. The Sertoli cell‐enriched preparation was obtained from seminiferous tubular fragments after sequential treatment with glycine buffer to remove peritubular‐myoid cells, a second collagenase digestion, mechanical fragmentation and washes to remove. germ cells. Purified Leydig cells were then cultured either alone or together with Sertoli cells in culture dishes coated with collagen, fibronectin and laminin in a chemically defined medium without serum. The influence of co‐culture on basal testosterone secretion was examined in 3 successive 48 h periods. During culture for 0–48 h, 48–96 h or 96–144 h, testosterone secretion (ng/106 Leydig cells, means f SEM) was: 9.7 ± 2.1, 6.1 ± 2.1, and 3.4 ± 1.3, by L2 Leydig cells; 81.6 ± 16.3, 82.6 ± 23.1, and 39.1 ± 13.6, by L3 Leydig cells; 19.0 ± 4.6, 44.0 ± 18.0, and 154 ± 69.3 by L2 + Sertoli cell co‐cultures; and 124.4 ± 13.2, 199.2 ± 14.7, and 346.1 ± 78.4, by L3 + Sertoli cell co‐cultures, respectively, showing that: (i) L3 Leydig cells secrete more testosterone than L2 Leydig cells, irrespective of whether they were cultured alone or with Sertoli cells; (ii) During all 3 time periods, co‐culture with Sertoli cells enhanced testosterone secretion by L2 and L3 Leydig cells; and (iii) during the course of culture, testosterone secretion decreased when Leydig cells were cultured alone and increased when Leydig cells were co‐cultured with Sertoli cells. When cultured in the presence of hCG (10–10M), testosterone secretion was enhanced during the first 48 h of culture, but not there after, in all types of culture. These data show a reproducible, time‐dependent, stimulatory effect of adult human Sertoli cells on basal testosterone secretion by adult human Leydig cells. It is concluded that culture of normal adult human Sertoli and Leydig cells can be performed with testicular tissue removed from young men, and in this in‐vitro system, Sertoli cells stimulate long‐term Leydig cell testosterone secretion. This study supports the concept of the control of Leydig cell function by Sertoli cells in the human testis.
Contents of fructose, citric acid, acid phosphatase, proteins and electrolytes in secretions of the accessory sex glands of the male golden hamster
Authors: Chow P. H. , Chan C. W. , Cheng L. Y. L.
Published: May 2008

Secretions were collected from the ampullary gland, dorsolateral prostate, ventral prostate, coagulating glands and seminal vesicles of male golden hamsters aged 15–20 weeks. The concentrations of total protein, citric acid, fructose, acid phosphatase, chloride, sodium, potassium, calcium, magnesium and zinc were determined. The ampullary gland secreted predominantly citric acid, sodium and acid phosphatase. Zinc was secreted only by the prostatic complex, with the largest quantity coming from the coagulating gland. The highest concentrations of potassium, calcium and magnesium were found in secretions of the ventral prostate. The coagulating gland and dorsolateral prostate were the principal contributors to total protein. The hamster appears to be more related to the mouse than to the rat in terms of the secretory functions of its accessory sex glands.
Immuno‐competent cells in the murine epididymis following infection with Escherichiu coli
Authors: Nashan D. , Jantos C. , Ahlers D. , Bergmann M. , Schiefer H. G. , Sorg C. , Nieschlag E.
Published: May 2008

Epididymitis was induced by retrograde injection of Escherichia coli into the vas deferens of 28 mice. A group of 28 saline‐injected animals served as controls. On Days 1, 3, 7 and 28, groups of seven animals were killed. Bacterial culture was performed. Leucocyte numbers and distribution were determined in epididymides.

In infected mice, E. coli were isolated from all epididymides on Days 1 and 3, but only from five of seven epididymides on Days 7 and 28. One week after infection, the total number of macrophages rose from about 10 to 28%. Significantly increased macrophage percentages were also found in animals killed 28 days after infection. A simultaneous increase in MHC class II positive cells was seen on Day 7. A total of 20% of the cells expressed MHC class II in infected epididymides (normal = 6%). A similar increase was found on Day 28 after infection. Most of the macrophages and MHC class II positive cells were located in the interstitium, fewer in the peritubular layer and nearly none in the epithelium. The main increase in these cells occurred in the interstitium and, to a lesser but significant extent, in the peritubular area.

T‐helper and T‐suppressor/cytotoxic lymphocytes reached peak values on Day 28. The increase in T‐lymphocytes and simultaneous appearance of plasma cells follo wed the increase in numbers of macrophages and MHC class II positive cells. They were located mainly in the interstitium. A sequential increase in leucocyte subsets and negative culture results for E. coli were observed on Days 7 and 28 (2/7 on each day). The inflammatory process was restricted to the interstitium.

It is concluded that stimulation of T‐lymphocytes and plasma cells by macrophages/MHC class II positive cells acts as an effective immune response to E. coli in the epididymal interstitiurn.
Analysis of spermatozoa from the proximal vas deferens of fertile men
Authors: WEN R.‐Q. , LI S.‐Q. , WANG C.‐X. , WANG Q.‐H. , LI Q.‐K. , FENG H.‐M. , JIANG Y.‐J. , ZHANG X.‐L.
Published: May 2008

Fluids from the left and right proximal vas deferens were collected from 105 normal fertile men by cannulating the vas deferens during vasectomy, and sperm parameters analysed. Sperm motility (73.1 k 13.3Y0), normal sperm morphology (75.2 k 11.1“/o), sperm viability (72.7 k 18.8%) and the hypo‐osmotic swelling test (73.3 k 19.2%) were in the normal range, compared with that of ejaculated spermatozoa. However, sperm Concentration in the proximal vas deferens (6274.6 k 5103.8 × 10” ml‐‘ was higher than that in semen. Sperm concentration in the right vas deferens was significantly higher (P<0.05) than that in the left and the percentage of spermatozoa showing abnormal cervical mucus penetration was Significantly higher (47%) for the left than for the right (18%). There were no anti‐sperm antibodies on the surface of spermatozoa from the vas deferens as determined by the sperm cervical mucus contact test and immuno bead test. These parameters of spermatozoa from the proximal vas may reflect those of spermatozoa from the human cauda epididymis.
Use of human zonae from cryopreserved oocytes in a test to assess the binding capacity of human spermatozoa
Authors: MORROLL D. R. , LIEBERMAN B. A. , MATSON P. L.
Published: May 2008

Aged unfertilized oocytes from an assisted conception programme were cryopreserved and then utilized after thawing in a zona‐binding assay. Oocytes frozen using dimethyl sulphoxide (DMSO) showed poor survival post‐thaw (2/40, 5%) compared to those frozen with propanediol (PROH) (63/134, 47%). When the zonae were exposed to spermatozoa from fertile donors, those frozen with DMSO showed a significantly higher number of bound spermatozoa than did those frozen with PROH (P< 0.002). In both groups, oocytes which failed to survive the freeze‐thaw processes had greater numbers of bound spermatozoa than did those which survived (P<0.05). Oocytes from cases of failed fertilization showed no difference in their rate of sperm binding compared with oocytes from cases in which some fertilization had occurred. Zonae frozen in PROH but which were from oocytes which were not viable after thawing were used to assess the binding of spermatozoa from men who had failed previously to fertilize their partner's oocytesin vitvoand spermatozoa from men with poor quality semen and who had elected for treatment by micro‐injection sperm transfer. The number of spermatozoa bound to zonae was reduced significantly in both groups compared to a fertile donor.
Beta 1 ‐integrins of spermatozoa: a flow cytophotometric analysis
Authors: GLANDER H.‐J. , SCHALLER J.
Published: May 2008

The expression of adhesion molecules on human ejaculated spermatozoa was investigated by means of flow cytophotometry using monoclonal antibodies against eight different adhesion molcules. Spermatozoa from infertility patients who exhibited normal semen parameters showed specific fluorescence after incubation with antibodies against very late antigens VLAα4, VLAα 5, VLAα 6, and corresponding matrix proteins laminin and fibronectin. Semen samples from men with teratozoospermia or oligoasthenoteratozoospermia were characterized by reduced expression of adhesion molecules. Normal and pathological semen samples differed mostly in the expression of VLAα5. After artificial induction of the acrosome reaction by a cold shock method and incubation with 2 mM Ca2+, a changed pattern of adhesion molecules on the sperm surface was found for VLAα6 as the most prominent molecule. After treatment of spermatozoa with 0.10% Triton X‐100, the number of cells with adhesion molecules increased significantly. This result suggests that adhesion molecules are an integral part of sperm structure.
Diurnal variations in scrotal temperature of normal men and patients with varicocele before and after treatment
Authors: LERCHL A. , KECK C. , SPITERI‐GRECH J. , NIESCHLAG E. , Bergh A.
Published: May 2008

Scrotal temperature was monitored using a portable data recorder for periods of 24 h in six normal volunteers and 48 infertile patients with unilateral varicocele while subjects pursued their regular daily activities. Temperatures during sleep (Ts) were generally higher than daytime values (TD), probably as a consequence of thermal insulation in bed. These diurnal variations were found to be less pronounced in the infertile patients than in volunteers (Ts‐ TD= 0.29°C ± 0.06°C us 0.88°C ± 0.12°C;P<0.01). Moreover, scrotal temperatures at night of patients and volunteers were indistinguishable statistically, but were different during daytime hours. After successful ligation or embolization of the spermatic vein in 16 patients, no change in scrotal temperature was observed. Although sperm counts were higher after treatment, this difference was not significant (67.4 ± 17.2 × 106vs 105.8 ± 25.5 ± 106; P>0.05). The data support the view that varicocele‐related damage to the testis results from a lack of adequate cooling, and that treatment does not normalize the temperature pattern.
Plasminogen activator activity and fertilizing ability of human spermatozoa
Authors: LISON D. , TAS S. , GENNART J.‐P. , PSALTI I. , de COOMAN S. , LAUWERYS R.
Published: May 2008

Mature spermatozoa contain a number of proteases that are supposed to contribute to their fertilizing ability. The present study was directed at plasminogen activator (PA), a protease that belongs to the group of serine proteases and converts the zymogen plasminogen to the active broad‐spectrum protease plasmin. To investigate the possible role of PA in the fertilization process, we have measured sperm‐bound PA activity in 63 patients included in an in‐vitro fertilization (IVF) programme and assessed their relationship to standard semen parameters and the rate of fertilization. PA activity was correlated significantly with the sperm count, as well as with sperm motility and morphology. Using logistic regression analysis, specific PA (pmol pNA 10‐‐6 cells minP2) was found to significantly influence the probability of fertilization. Other significantly predictive factors were motility and the percentage of spermatozoa with normal morphology. The sperm concentration (106 cells ml‐1) did not significantly affect the outcome of IVF. We suggest that sperm‐bound PA is involved in the fertilization process and may represent a potential indicator of sperm fertilizing capacity.
The detection of antisperm antibodies in serum: a comparison of the tray agglutination test, indirect immunobead test and indirect Spermcheck assay
Authors: MORROLL D. R. , LIEBERMAN B. A. , MATSON P. L.
Published: May 2008

Testing for antisperm antibodies (ASAs) is an important part of the work‐up of the sub‐fertile couple, yet there is little consensus regarding the most appropriate methods. The Spermcheck assay (GSC; Bio‐Rad Laboratories Inc., Diagnostics Division, Hercules, CA, U.S.A.) is supplied with wash buffer, controls and bead reagent which detects all three major classes of ASAs (IgA, IgG and IgM) in a single test. This study compared results on a bank of samples using the tray agglutination test (TAT), immunobead test (IBT), GSC and a modified Spermcheck assay to detect a single isotype in each test (SISC). The IBT and SISC showed excellent correlation, with 127/141 (90.1%) tests agreeing. There was an apparent lack of sensitivity to IgM with GSC as 8/15 (53.3%) samples testing positive with IBT and 7/15 (46.7%) testing positive with SISC were negative with GSC. Of the 24 IBT‐negatives, seven (29.2%) were positive for TAT, indicating a high incidence of non‐immunological agglutination, though this decreased as the TAT titre increased. The proportion of samples testing positive for IBT increased with TAT titre: 3/20 (15.0%) for TAT‐negative samples, 6/10 (60.0%) for low titres and 21/24 (87.5%) for high titres. This was also observed when comparing the GSC with TAT. The TAT therefore appears useful as a first‐line screen, whilst the inability of the GSC to adequately detect IgM limits its use as an indirect test. Both the IBT and SISC can be used to further investigate the type and class of ASA present.
Immunohistochemical localization of calmodulin in the testes of patients with idiopathic male infertility
Authors: TSUJI Y. , SANO M. , NAGAHAMA M. , TSUTSUI Y. , YAMAMOTO M. , MIYAKE K.
Published: May 2008

The localization of calmodulin in testes of patients with idiopathic male infertility was studied using the indirect immunoperoxidase method. Specimens were obtained by testicular biopsy from 55 patients. They were divided into 26 cases of hypospermatogenesis, 11 cases of maturation arrest (8 of primary spermatocyte arrest and 3 of spermatid arrest) and 18 cases of Sertoli cell‐only syndrome. Regardless of the type of testicular pathology, the types of immuno‐reactive cell and the intensities of staining were the same as those in the normal testis. That is, staining for calmodulin was first found to be positive in early pachytene primary spermatocytes. It became intense in late pachytene primary spermatocytes and round spermatids. By contrast, elongated spermatids and spermatozoa were not stained. Sertoli cells were stained slightly or not at all. A calmodulin‐staining index (CaM‐S index) was defined as the proportion of primary spermatocytes that were stained intensely for calmodulin relative to the total number of primary spermatocytes. The indices for the testes of men with complete spermatocyte maturation arrest were significantly lower than those for the testes of normal controls and of men with hypospermatogenesis.

Degenerating late pachytene spermatocytes observed in the testes of men with spermatocyte arrest showed low calmodulin‐specific immunoreactivity. Such a decrease in numbers of normal late pachytene spermatocytes might be responsible for the low CaM‐S index in cases of complete spermatocyte arrest.
Arachidonic acid metabolites as intratesticular factors controlling androgen production
Authors: REDDY G. PANDURANGA , PRASAD M. , SAILESH S. , KUMAR Y. V. KIRAN , REDDANNA P.
Published: May 2008

The effect of inhibitors and products of arachidonic acid metabolism on rat testicular steroidogenesis has been investigated. In the presence of indomethacin (inhibitor of cyclooxygenase) and nordihydroguaiaretic acid (NDGA) (inhibitor of lipoxygenase), the activity of 3P‐hydroxy steroid dehydrogenase (3β‐HSD) and 17β‐hydroxysteroid dehydrogenase (17β‐HSD) were both inhibited. The LH‐stimulated increase in secretion of testosterone and progesterone was also inhibited by indomethacin and NDGA. On the other hand, vitamin E (anti‐oxidant and inhibitor of lipoxygenase), stimulated the activity of both 3β‐HSD and 17β‐HSD and enhanced LH‐stimulated androgen production. The metabolites of lipoxygenase (15‐HPETE, 15‐HETE, 5‐HPETE and 5‐HETE) and cyclooxygenase (PGF2α) pathways stimulated 3β‐HSD and 17β‐HSD activity and enhanced the secretion of progesterone and testosterone. It is concluded that arachidonic acid matabolites are intratesticular factors which can regulate LH‐stimulated testicular steroidogenesis.
Experimental testicular germ cell tumorogenesis in mouse strains with and without spontaneous tumours differs from development of germ cell tumours of the adult human testis
Authors: Walt H. , Oosterhuis J. W. , Stevens L. C.
Published: May 2008

The aim of this study was to undertake a morphological analysis of the earliest stages of experimentally induced (by genital ridge grafting) germ cell tumours in mouse strains with (129/Sv‐ter) and without (MA) spontaneous tumorogenesis. Genital ridges from fetuses aged 12 or 13 days from 129/Sv‐ter and MA were transplanted into the testes of adult 129/Sv‐ter. The results show clearly that experimentally induced carcinoma‐in‐situ in mouse testes differs considerably from its human counterpart, found in patients with and without testicular germ cell tumours, and considered to be the precursor for all kinds of germ cell tumours of the adult testis apart from spermatocytic seminoma. The results indicate that development of testicular germ cell tumours is different in man and the mouse.
Evaluation of possible determinants and consequences of Leydig cell heterogeneity in man
Authors: QURESHI S. J. , SHARPE R. M.
Published: May 2008

Leydig cells in the human testis are highly heterogeneous, consisting of variably staining light and dark cells. The basis for this difference is unknown. The present study has assessed whether differing numbers or proportions of dark and light Leydig cells are related: (1) to the pronounced inter‐individual variation in testosterone production by isolated Leydig cells, and (2) to differences in structural composition of the testis. Testes (paired weight 6.6–59.48 g) were obtained from 27 men aged 72.9 ± 9.5 years (range 54–89 years) undergoing orchidectomy as primary treatment for prostatic cancer. Leydig cells were isolated by Percoll‐purification and cultured for 20 h under basal and hCG‐stimulated conditions. The proportion of light and dark Leydig cells isolated by this method was shown to reflect their proportions in situ, based on the morphometric analysis of fixed testicular tissue from the same men. Leydig cells isolated from all testes produced testosterone in vitro and responded to stimulation by hCG, though the amounts of testosterone produced varied widely between subjects. Because of the latter, samples were grouped into ‘low’ (n= 9), ‘medium’ (n = 11) and ‘high’ (n= 7) groups on the basis of their testosterone production. These groups did not differ in their age, testicular size or gross testicular morphology, though men in the ‘high’ group tended to have more total Leydig cells per testis. However, there was no overall correlation between testosterone production by isolated Leydig cells and the numbers of light or dark Leydig cells or their ratio or the total number of Leydig cells per testis. The relationship between the volume of light and dark Leydig cells and testicular composition was also assessed. The volume of both types of Leydig cells was strongly correlated (p <0.001) with the volume of germ cells, but otherwise light and dark Leydig cells correlated positively with different structures. Thus, the volume of light Leydig cells correlated (p < 0.001) with the volume of blood vessels and of peritubular tissue whereas the volume of dark Leydig cells correlated (p < 0.01) with that of the tubular lumen. These differences could indicate differences in regulation and/or function of light and dark Leydig cells. However, the present data do not support the idea that light and dark Leydig cells may differ in their steroidogenic capacity.
Successful gonadotrophin treatment of hypogonadism in postoperative patients with macroprolactinoma and persistent hyperprolactinaemia
Authors: ISURUGI K. , KAJIWARA T. , HOSAKA Y. , MINOWADA S.
Published: May 2008

We report on two males with prolactinoma in whom hyperprolactinaemia and hypogonadism persisted for several years postoperatively despite the administration of a dopamine agonist or bromocriptine. In these patients, a GnRH test revealed no response in the levels of serum LH or FSH. An hCG stimulation test provoked no response in the serum levels of testosterone. Case 1, who was 28 years old at the first visit, received parenteral testosterone and appreciable virilization of the genitalia was noted within a few months. When he married and desired to father a child, the treatment was switched to hCG/hMG combined therapy and spermatozoa appeared subsequently in the ejaculate, although their numbers were low. His wife conceived and delivered a healthy baby girl. Case 2 was a single young man who presented with hypogonado‐trophic hypogonadism and hyperprolactinaemia. He was started on hCG injections three times per week and the maturation of his genitalia was advanced rapidly. Semen analyses showed sperm concentration and motility to be within the normal range. Post‐treatment GnRH test revealed no improvement in gonadotrophin responses for LH or FSH. In both cases, the hCG test repeated after the gonadotrophin treatment showed normal basal and stimulated testosterone levels. During the course of gonadotrophin treatment in these cases, serum prolactin levels remained elevated, and it is suggested that, in the two cases, the hypothalamo‐pituitary function was disturbed by the tumour or its manipulation and the capacity of the pituitary gland to secrete gonadotrophin was impaired. Under such circumstances with persisting hyperprolactinaemia, hCG and/or hCG/hMG combination treatment can induce normal virilization and advance spermatogenesis sufficiently to achieve fertility.
Enzymohistochemical and immunohistochemical study of the human efferent ducts
Authors: REGADERA J. , PALACIOS J. , MARTÍN‐CÓRDOVA C. , NISTAL M. , COBO P. , PANIAGUA R.
Published: May 2008

An enzymohistochemical and immunohistochemical study of the efferent ducts was performed in normal adult men. The epithelium donsists of two types of columnar cells: principal cells (PCs) and ciliated cells (CCs), and is surrounded by a lamina propria (LP) with cells arranged circularly (LPCs). Enzymohistochemical study revealed more intense activity of succinic dehydrogenase, NADP, and ATPase in the CCs than in the PCs. The LPCs also showed an intense reaction for NADP and ATPase. Acid phosphatase activity was only intense in the apical cytoplasm of PCs. Immunohistochemical study revealed that antibodies to oestradiol receptor‐related protein (ER‐D5) immunostained the PCs and CCs intensely and the LPCs weakly. AEl/AE3 antibodies (which stain keratins nos. 1–8 and14, 15 and 19) immunostained the PCs intensely, but was negative in both CCs and LPCs. Antibodies to keratin Ks.4.62 (which stain keratin no. 19) immunostained PCs and CCs but not LPCs. Epithelial membrane antigen antibodies (EMA) immunostained the adluminal surface and apical cytoplasm of PCs. Anti‐vimentin antibodies immunostained the cytoplasm of PCs and CCs weakly as well as isolated cells in the LP. Antibodies to desmin immunostained most LPCs. Antibodies to collagen IV immunostained the basal lamina and many extracellular spaces in the LP, mainly around the LPCs. The differences between the enzymohistochemical and immunohistochemical patterns of the efferent ducts and those of the epididymis may help to explain functional differences along the epididymis.
Sentinels of Leydig cell structural and functional changes in golden hamsters in early testicular regression and recrudescence
Authors: HIKIM A. P. SINHA , CHANDRASHEKAR V. , BARTKE A. , RUSSELL L. D.
Published: May 2008

The seasonally breeding hamster model was used to assess changes associated with Leydig cell activity and inactivity. Specifically, parameters of Leydig cell structure and function were studied to determine the early changes seen in seasonally breeding golden hamsters in photoperiod‐induced gonadal regression and photoperiod‐induced gonadal recrudescence. Time intervals used to characterize early regression and recrudescence were selected with the objective of determining the most sensitive parameters characterizing functional transitional states. In early regression, plasma levels of follicle‐stimulating hormone (FSH) but not luteinizing hormone (LH) or testosterone were reduced significantly. Regressive structural changes included decreases in volume of the interstitium, total number of Leydig cells, blood vessels, and the seminiferous tubules and tubular lumen. A decrease in volume, but not numbers of Leydig cells, was accompanied by decreases in volume of Leydig cell tubular smooth endoplasmic reticulum (t‐ER), Golgi complex, and the peroxisomes and decreases in surface area of the inner mitochondrial membrane and t‐ER, suggesting that early Leydig cell changes are restricted primarily to organelles associated with steroid biosynthesis. In early recrudescence, at a time when there was an increase in the number of germ cells in the basal compartment, blood levels of LH, FSH and testosterone were all increased. There were increases in testicular weight, volume, seminiferous tubular lumen, blood vessel and interstitial volumes. Leydig cells increased in size as a result of increases in nuclear, nucleolar and cytoplasmic volumes, while Leydig cell numbers did not increase. At the subcellular level there were increases in the surface areas of the cell, mitochondrial membranes and cisternal endoplasmic reticulum (sparsely populated with ribosomes). Unlike the changes seen in early degeneration when steroid synthetic organelles were initially affected, the changes in early recrudescence indicate that Leydig cells must first rebuild their synthetic machinery (nucleus, nucleolus and rough endoplasmic reticulum) that, at a later time, will reconstitute the cell's steroidogenic machinery. Correlation of hormonal parameters with structural parameters did indicate a relationship between hormonal parameters and steroid secreting organelles. Correlations were strongest with testosterone but, surprisingly, plasma FSH levels correlated more strongly with many structural parameters of the Leydig cell than did the levels of LH. Since FSH receptors are not present on Leydig cells, these data add to the growing data suggesting a role for factors originating from the seminiferous tubule in modulation of Leydig cell function.
The Gubernaculum in adult female, adult male and TFM male mice
Authors: GRIFFITHS A. L. , MOMOSE Y. , HUTSON J. M.
Published: May 2008

There is still controversy over whether androgens are involved in gubernacular migration during descent of the testis, prompting a review of anatomical events controlled by androgens. The gubernacular position in adult female, male, and testicular feminized male (TFM) mice (with complete androgen resistance) was observed macroscopically to determine its caudal limit and whether the adult TFM male still contained guvernacular bulk. The length of the processus vaginalis was measured, and the presence of a differentiated cremaster muscle was determined histologically. In 10 adult female mice there was neither a processus vaginalis nor a cremaster muscle, and the gubernaculum (round ligament of the ovary) attached to the external inguinal ring. In 10 adult TFM mice the gubernaculum ended at or just beyond the external ring, and was present in an amount corresponding to its size at the end of the outgrowth phase of development, with secondary infiltration by fat. The processus vaginalis was poorly developed, the cremaster muscle was undifferentiated, and there was no eversion of the cremaster sac. In 10 normal adult male mice the gubernaculum and testis had migrated to the scrotum, and the gubernaculum had regressed fully. Because gubernacular development in the androgen‐resistant mouse ceases after termination of transabdominal testicular migration, the anatomy of the adult confirms that, in the mouse, normal androgenic function is essential for eversion of the cremaster sac and gubernacular migration beyond the groin to the genital folds.
Collective isolation of two major, close molecular weight coagulum proteins occurring in human semen before liquefaction
Authors: MANDAL ARABINDA , BHATTACHARYYA ASOK K.
Published: May 2008

Two major, close molecular weight basic coagulum proteins of freshly ejaculated human semen identifiable on reducing SDS‐PAGE, have been isolated together from washed seminal coagulum. The latter was solubilized in urea, followed by reduction and carboxymethylation of disulphide bonds. By high‐resolution chromatography on carboxymethy‐Sephadex and Sephacryl S‐300 using 8 M urea in 0. 1m Tris‐HCl (pH 7.4) as eluate these proteins were isolated together (purity 99%) with a yield of 10.5%. The molecular weights of the isolated proteins on SDS‐PAGE were 75 and 79 kD.
Management of bilateral nonpalpable testes: laparoscopic diagnosis and orchidectomy
Authors: HAUSER R. , LESSING J. B. , SAMUEL D. , YAVETZ H. , PEYSER M. R. , PAZ G. F. , HOMONNAI Z.
Published: May 2008

Bilateral nonpalpable testes in the adult human are associated with testicular malignancy, infertility and other abnormalities. Investigation for localization of the testes is mandatory, and either orchidopexy or orchidectomy is indicated. Laparoscopy was performed in an azoospermic male with bilateral unpalpable testes. A diagnosis of intra‐abdominal testes was made and bilateral orchidectomy was performed. Laparoscopy is recommended for diagnosis and precise localization, as well as for orchidectomy, thus avoiding open abdominal operation in cases of intra‐abdominal testes.
Expression of the c‐kit protein product in carcinoma‐in‐situ and invasive testicular germ cell tumours
Authors: MEYTS E. RAJPERT‐DE , SKAKKEBKK N. E.
Published: May 2008

Carcinoma‐in‐situ of the testis (CIS) is the precursor of invasive germ cell tumours. It is believed that CIS cells may originate from early fetal gonocytes. Recently, the proto‐oncogene c‐kit has been implicated as crucial for the development and migration of primordial germ cells. In this study, CIS and overtly invasive human male germ cell tumours were analysed immunohistochemically for expression of the c‐kit proto‐oncogene protein product. Testicular tissue samples from 36 patients with various types of testicular germ cell neoplasia and 19 control specimens were stained using an indirect immunoperoxidase method. High expression of c‐kit was found in almost all cases of CIS, both when the lesion was the only pathology, and when CIS was adjacent to invasive tumours. The Kit staining was retained in seminomas with variable intensity; the majority of cells in tumour mass exhibited c‐kit expression in 61% of the samples while focal expression was observed in 39% of the samples studied. No expression of c‐kit was detected in non‐seminomas or in normal testicular germ cells. High expression of the proto‐oncogene in CIS cells supports the hypothesis of their origin from primordial germ cells. In addition, we propose that the c‐kit protein product is a new marker for carcinoma‐in‐situ of the testis.
Seminal sperm antibodies exhibit an unstable spontaneous course and an increased incidence of leucocytospermia
Authors: PASCHKE R. , BERTELSBECK D. SCHULZE , BAHRS S. , HEINECKE A. , BEHRE H. M.
Published: May 2008

The prognostic significance of seminal sperm antibodies for male fertility is difficult to define. Among other factors, spontaneous remissions and transient induction through genito‐urinary infections may change their significance for male fertility considerably. We therefore investigated their spontaneous course over time and their association with leucocytospermia. For the investigation of possible relationships between the mixed antiglobulin reaction (MAR) test results for IgA and IgG sperm antibodies and leucocytospermia, 138 patients with <1 million leucocytes/ml ejaculate were compared with 1051 patients with <1 million leucocytes/ml ejaculate (WHO normal range). In a second part of the study the spontaneous course of MAR IgG and IgA test results was investigated in 58 of the 1189 patients who had three or more MAR tests and a leucocyte concentration of <1 million/ml ejaculate. The mean interval between the first and second MAR test was 4.1 (SD ±4.2) and between the second and third MAR test 6.0 (SD ± 6.7) months. In total, 64% of patients with an MAR IgG test result >40% showed a stable spontaneous course. Patients with lower MAR IgG test results and the majority of all MAR IgA‐positive patients were found to have an undulating course of MAR test results. There was a higher incidence of leucocytes <1 million/ml ejaculate in patients with higher MAR results (p <0.05 for IgG, p <0.001 for IgA). The unstable spontaneous course of MAR IgG results <40% and all MAR IgA results suggests that evaluations of the in‐vivo significance of seminal sperm antibodies for male fertility should be based on repeated MAR determinations. Moreover, the association of leucocytospermia (<1 million leucocytes/ml ejaculate) with an increased incidence of positive MAR test results suggests a thorough evaluation, treatment and follow‐up of possible genital tract infections in MAR‐positive patients with <1 million leucocytes/ml ejaculate.
Analysis of spermatozoa from the proximal vas deferens of vasectomized men
Authors: WEN R‐Q. , LI S‐Q. , WANG C‐X. , WANG Q‐H. , LI Q‐K. , FENG H‐M. , JIANG Y‐J. , HUANG J‐C.
Published: May 2008

This study assessed the condition of spermatozoa from the proximal vas deferens of men after vasectomy. The fluids of both proximal vas deferens were collected from 67 vasectomized men by cannulating the vas deferens at the time of vasectomy reversal. Selected sperm parameters were analysed after incubation of the spermatozoa for 30 min at 37°C. Spera concentration in the proximal vas from vasectomized men (16 312 ± 21 496 million per ml, geometric mean: 7948 ± 398 million per ml) was significantly higher than that of fertile men and was maintained at a constant level independent of the duration of vas obstruction. The means of sperm motility (36.2 ± 26.2%), spermatozoa with normal morphology (50.7 ± 21.7%), sperm viability (53.0 ± 25.3%) and hypo‐osmotic swelling test (HOS‐test, 53.9 ± 21.7%) were statistically lower than the respective values for normal fertile men. There was no significant correlation between the duration of vas obstruction and the above semen parameters. In 46.4% of vas fluids all spermatozoa were immotile and this condition was more common after 3 years of vasectomy. Immotile spermatozoa in the proximal vas fluids at the time of vasectomy reversal may be an important factor for predicting semen quality and fertilizing ability after vasovasostomy. There were no significant differences in the results of sperm‐cervical mucus penetration test (CMPT) between spermatozoa fiom vasectomized and fertile men. Antisperm antibodies on the surface of spermatozoa from the vas of vasectomized men were determined by the immunobead test (IBT; 78.6% for IgG, 32.1% for IgA) and sperm cervical mucus contact test (SCMC, 36.4%). The presence of antisperm antibodies on the spermatozoa from the vas of vasectomized men may explain, in part, the lower pregnancy rate after vasovasostomy. These parameters of spermatozoa from the proximal vas of vasectomized men may closely reflect those in the cauda epididymis after vasectomy.
Assessment of protein synthesis and secretion by rat seminiferous and epididymal tubules in vivo
Authors: TURNER T. T. , AVERY E. A. , SAWCHUK T. J.
Published: May 2008

In vivo microperifbsion and micropuncture were used to study tubule protein synthesis and proluminal secretion by the male reproductive tract in vivo. Somniferous and caput and cauda epididymal tubules were perihsed for 3 h. with [35S]‐methionine. Perifused interstitial fluid (IF), lumen fluid (LF), and tubule extract (TE) were collected. Proteins were separated by SDS‐PAGE, and autoradiograms were developed.

Trichloroacetic acid precipitable proteins in each fluid were determined and a protein synthesis index (PSI) was calculated. PSI values demonstrated that the cauda epididymis synthesized less protein in vivo than did either seminiferous or caput tubules.

Seminiferous tubules synthesized and secreted into the tubule lumen a relatively constant panel of proteins. Epididymal tubules synthesized and secreted proteins in a region‐specific manner. In the caput epididymis the most prominent secreted bands were consistent with the heavy and light chains of epididymal clusterin. In the cauda epididymis, the most prominent synthesized and secreted protein was a 25 kDa protein consistent with the protein D. The above approach to studying protein synthesis and secretion will allow direct study of the physiological and path physiological effects on this important epithelial hnction in vim
Influence of hypothyroidism induced at prepuberty on epididymal lipids and the number and motility of spermatozoa in rats*
Authors: KUMAR P. N. S. , ARULDHAS M. M. , JUNEJA S. C.
Published: May 2008

Fifteen prepubertal male rats (age 30 days) were divided into three groups of five each: Group 1, hypothyroid (Tx)—rats were thyroidectomized at day 30; Group 2, T4 &‐thyroxine) replacement therapy (Tx+T4)—rats were thyroidectomized at day 30 and treated daily i.m. with T4 (6μg/100 g body weightlday) for 30 days from day 31 to day 60 post‐thyroidectomy (age 90 days); Group 3, control—rats were sham‐operated and treated with vehicle. The rats from all groups were killed on day 61 post‐thyroidectomy or post‐sham operation (age 91 days). The serum levels of testosterone, T, and T, decreased in the Tx group (p<0.001). In the Tx+T4 group the levels of T, and T, were restored to control values, whereas testosterone levels remained lower than in the control group. Hypothyroidism caused various changes in the levels of epididymal phospholipids and neutral lipids. These were restored differentially or were altered further in the Tx+T4 group. The number and forward motility of spermatozoa, recovered from the cauda epididymis, were decreased significantly (p<0.01) in the Tx group and were not restored in the Tx+T4 group. This study shows that chronic hypothyroidism, induced at prepuberty and continued for 60 days, causes various changes in the lipid composition of the caput and cauda epidldymis and lowers the quality and quantity of spermatozoa in the cauda epididymis. T4 therapy for 30 days, especially during the postpubertal period, did not restore the quality or quantity of spermatozoa and caused differential changes in the levels of epididymal lipids, depending upon the region. The study may be helpful in revealing the reasons for subfertility in hypothyroid male patients.
An in vitro promoting role for hydrogen peroxide in human sperm capacitation
Authors: GRIVEAU J. E. , RENARD P. , LANNOU D. Le
Published: May 2008

A complex process of maturation called capacitation is an essential step for spermatozoa to fertilize oocytes. Recent studies have shown that reactive oxygen species (ROS) can enhance the capacitation of human spermatozoa and sperm‐zona interaction. We have investigated whether hydrogen peroxide (H2O2) could trigger capacitation of human spermatozoa and the acrosome reaction. The addition of catalase, a specific H202 scavenger, at the beginning of the capacitation process decreased the levels of both hyperactivation and induced‐acrosome reaction whereas catalase added 15 min before the induction of the acrosome reaction by the calcium ionophore had no effect. Supplementation of the medium with H2O2 resulted in increased levels of hyperactivation and the acrosome reaction, whereas H2O2 added 15 min before induction of the acrosome reaction did not have any stimulatory effect. These results suggest that H2O2 may be involved in the capacitation process of human spermatozoa but not in the acrosome reaction.
Effect of pentoxifylline on experimentally induced lipid peroxidation in human spermatozoa
Authors: GAVELLA M. , LIPOVAC V.
Published: May 2008

We demonstrated previously that pentoxifylline in millimolar concentrations can inhibit superoxide anion production by human spermatozoa. In the present study we have examined the effects of the same concentrations of pentoxifjrlline on experimentally induced lipid peroxidation, as measured by malondialdehyde formation in the thiobarbituric (TBA) assay. Under the experimental conditions used, preincubation of spermatozoa with pentoxifjdline led to a significant dose‐dependent stimulation (p<0.005) of malondialdehyde production amounting to 10.77 ± 2.35%, 13.45 ±2.99% and 17.4 ± 1.99% (mean ± SEM) for 1.9, 3.7 and 11.2 mmol/l pentoxifylline, respectively. In the presence of 11.2 mmol/l pentoxifylline, an increase in iron‐catalysed lipid peroxidation potential was detected in samples of spermatozoa from 29 infertile men, regardless of their initial levels of malondialdehyde. The results of this study indicate that pentoxifylline might further augment the ferrous ion‐stimulated decomposition of pre‐accumulation lipid hydroperoxides in the sperm plasma membrane and thus promote malondialdehyde generation in the TBA assay.

It is concluded that the stimulatory effect of pentoxifjdline on iron‐induced lipid peroxidation may have an adverse effect on the quality of sperm suspensions prepared for in vitro fertilization, a possibility which should be investigated further.