bluuebunny/crossref_metadata_2025 huggingface dataset:
Developmental pattern of androgen‐regulated proteins in seminal vesicles from the mouse
Authors: NORMAND T. , JEAN‐FAUCHER CHRISTIANE , JEAN C.
Published: May 2008

Proteins from secretions or homogenates of mice seminal vesicles were analysed by polyacrylamide gel electrophoresis. In homogenates about 15 bands were differentially induced with molecular weights (MW's) of 12, 13. 14, 15, 15.5, 71, 120 and 140 kD, or repressed molecular weights of 12.5, 14.3, 28, 30, 53, 73, 90–105 kD). The effects of castration were reversed by testosterone and dihydrotestosterone but not by oestradiol, progesterone or corticosterone. When the androgen‐dependence of proteins was investigated using radioactive methionine the protein spectra showed that about 12 bands with molecular weights of 13, 13.7, 14, 15, 15.5, 16, 20.5, 24, 37, 38.5, 56, 68, 96 and 180 kD were differentially induced or repressed by androgens. Of the induced proteins, those with low molecular weight (12–15.5 kD) were accumulated in significant amounts between 20 days and 30 days, coincident with the pubertal increase of androgens in the seminal vesicles. Those induced proteins with high molecular weight (71, 120 and 140 kD) appeared between 40 days and 60 days. The androgen‐repressed proteins were strongly evident in immature males, but disappeared after day 40.
Synthesis of extracellular matrix components by somatic testicular cells from immature and pubertal rats
Authors: RODRÍGUEZ J. P. , MINGUELL J. J.
Published: May 2008

Total testicular cells derived from immature and pubertal rats were cultured under long‐term conditions. Somatic adherent cells proliferated in culture and produced collagen and proteoglycans. Collagen synthesis accounted for 25% and 5% of total protein synthesized by adherent cells derived from immature, and pubertal rats, respectively. Proteoglycan synthesis was higher in cells from immature than from pubertal rats. The proportion of different types of glycosaminoglycan chains (particularly hyaluronic acid and chondroitin sulphate) also varied according to the age of the donor. The results suggest that the synthesis of extracellular matrix components by somatic testicular cells is an age‐related process which probably plays an active role in spermatogenesis.
Antisperm antibodies and sperm motility: a study using timed exposure photomicrography
Authors: ADEGHE A. J.‐H. , ZHANG J. , CUTHBERT J. , OBHRAI M.
Published: May 2008

This study examined the effect of antisperm antibodies on sperm motility. Anti‐sperm antibodies present in seminal plasmas with different sperm agglutinating titres were transferred passively to normal donor sperm, and the effects on movement characteristics (velocity of forward progression, amplitude of lateral head displacement, percentage progressive motility and percentage non‐progressive motility) were analysed using timed exposure photomicrography. There was no significant association between sperm movement characteristics and the presence or titre of antisperm antibodies in seminal plasma. Furthermore, no differences were detected between those samples that possessed sperm agglutinating versus sperm immobilizing activity. These findings do not support the common belief that antisperm antibodies are a cause of poor sperm motility in semen.
Does computerized image analysis of sperm movement enhance the predictive value of semen analysis for in‐vitro fertilization results?
Authors: GRUNERT J.‐H. , GEYTER C. DE , BORDT J. , SCHNEIDER H. P. G. , NIESCHLAGM E.
Published: May 2008

The influence of semen quality on fertilization rates in an in‐vitro fertilization (IVF) programme was studied by analysing both conventional semen parameters and computerized movement characteristics. The study was based on 407 inseminated oocytes which were obtained from 50 patients in 113 laparoscopies. Sperm concentration did not correlate strongly with the fertilization rate. Sperm motility and morphology were the most meaningful parameters in predicting fertilization success. A drop in fertilization rate was found when sperm motility or normal morphology were below 40%. Sperm velocity measured in semen was the only sperm movement parameter which correlated with the fertilization rate, albeit weakly. The latter was reduced when average sperm velocity in semen was < 50 μm/sec. Conventional semen parameters seem to be more predictive of the fertilizing potential of an ejaculate than movement characteristics obtained by computerized image analysis.
Absence of correlation between the levels of ATP and other seminal compounds in semen and the results of human in‐vitro fertilization
Authors: MIEUSSET R. , PARINAUD J. , CHAP H. , PONTONNIER G.
Published: May 2008

As part of an in‐vitro fertilization programme (IVF), semen levels of adenosine triphosphate, fructose, citrate, zinc, free L‐carnitine and glycerophosphocholine were measured in 178 men. None of these substances appeared to have any predictive value with regard to the fertilizing potential of the semen during IVF.
Quantitative evaluation of the maintenance and development of spermatocytes and round spermatids in cultured tubule fragments from immature rat testis
Authors: TOEBOSCH A. M. W. , BRUSSÉE R. , VERKERK A. , GROOTEGOED J. A.
Published: May 2008

Maintenance and development of spermatocytes and round spermatids was studied in an in‐vitro incubation system. This system consisted of open tubule fragments from 26–day‐old rat testes, obtained after collagenase treatment. The tubule fragments contained Sertoli cells and spermatogenic cells up to and including a small number of early round spermatids. The number of primary spermatocytes and round spermatids in the tubule fragments was estimated using flow‐cytometric analysis, immediately after isolation and after 72 h of incubation. In addition, the activity of LDH‐C4 in the tubule fragments was measured. After 72 h of incubation, the percentage of spermatocytes was reduced by 70–80%, but the percentage of spermatids was doubled. The total LDH‐C4 activity per well was increased 2–3–fold during 72 h of incubation of the fragments. A modest improvement of the culture results was observed when a combination of FSH, insulin, retinol and testosterone was added to the medium. LDH‐C4 activity was investigated to see whether it could be used as a quantitative marker of isolated and cultured spermatocytes and spermatids. It was observed that LDH‐C4 activity per cell was decreased when spermatocytes and spermatids were isolated and/or incubated at 4°C. However, the cellular enzyme activity returned to control values during subsequent incubation of the cells at 32°C, either in the absence or presence of a protein synthesis inhibitor. Cellular LDH‐C4 activity may be influenced not only by temperature, but possibly also by other cell isolation conditions. It is concluded that LDH‐C4 activity may not be a reliable quantitative marker for the presence of spermatocytes and spermatids in culture, but should be used in combination with other analytical methods such as DNA estimation and DNA flow cytometry.
Variation in, and inter‐relationship between, prostaglandin levels and other semen parameters in normal men
Authors: SVANBORG K. , GOTTLIEB C. , BENDVOLD E. , YGDEMAN M. B.
Published: May 2008

In each of 135 ejaculates from 46 healthy men, semen volume, various sperm parameters and the level of prostaglandins (PGs) were determined. Data were analysed to determine the within‐ and between‐subject variability, the effect of abstinence period and the correlation between different parameters. Data which did not reveal a Gaussian distribution were transformed logarithmically. For all variables the between‐subject variance exceeded the variation within subjects (P < 0.001). Variation in time of abstinence between 1 and 3 days influenced semen volume and sperm content as well as the content of 19‐hydroxylated PGs, but did not affect sperm motility or the concentration of PGE or PGF. A significant negative correlation was found between the seminal content of PGE and PGF and the total sperm count as well as the sperm concentration. Sperm motility was related to the balance between the content of 19‐hydroxy PGE and 19‐hydroxy PGF but not to their separate concentrations. The ratio between 19‐hydroxy PGE and 19‐hydroxy PGF did not change when the time of abstinence differed. Synthesis of the two 19‐hydroxylated PGs seemed to be regulated by the same mechanism, but their hydroxylation capacity still varied considerably between individuals.
Purification of an acrosomal antigen recognized by a monoclonal antibody and antifertility effects of isoimmune serum
Authors: LIU M.S. , YANG Y. , PAN J. , LIU H.‐W. , MENGE A. C. , LEE C.‐Y. G.
Published: May 2008

A highly conserved acrosomal antigen reactive to a monoclonal antibody (HS‐63), generated against human sperm, was purified to homogeneity with a combination of conventional procedures and immunoaffinity chromatography using a soluble extract of mouse and rabbit testes. The molecular weight of the purified antigen was 42–50 kD when analysed by sodium dodecylsulphate polyacrylamide gel electrophoresis. The high specificity of the purified antigen to monoclonal antibody HS‐63 was shown by indirect immunofluorescent inhibition assay, enzyme‐linked immunosorbent assay, Western blot analysis and radioimmunosorbent assay. The purified antigen was used for isoimmunization of mice and rabbits. Following successive immunizations, antisera of high titres were raised and reacted specifically with antigen on the sperm acrosome and in testes of several mammalian species, but not with somatic tissues. These isoimmune sera exhibited strong inhibition on mouse in‐vitro fertilization and human sperm penetration of zona‐free hamster eggs. The results of this study suggest that the sperm‐specific acrosomal antigen reacting with HS‐63 could be a good candidate for the development of immunocontraceptive vaccines in humans and in other animals.
Presence and distribution of leucocyte subsets in the murine epididymis after vasectomy
Authors: NASHAN D. , COOPER T. G. , KNUTH U. A. , SCHUBEUS P. , SORG C. , NIESCHLAG E.
Published: May 2008

Male mice were vasectomized by ‘open‐ended’ or ‘closed’ techniques. After 4 weeks the cell‐mediated immune reactions were compared with those of sham‐operated animals by immunohistochemical localization of leucocytes, using specific monoclonal antibodies. Macrophages and MHC class II antigen‐positive cells were the major cell types to appear in all regions of the epididymis after both types of operation. There was recruitment of T‐helper/inducer leucocytes but not of T‐suppressor‐cytotoxic cells. An increased presentation of macrophage‐migration inhibiting factor antigen appeared in interstitial and peritubular locations. After ‘closed’ and ‘open‐ended’ vasectomy granulomata developed in the epididymis. The sperm‐containing lumen of these granulomata was invaded by macrophages, MHC class II‐positive cells and T‐helper/inducer lymphocytes. This mouse model thus reveals a significant epididymal inflammatory response of the epididymis to vasectomy.
Laminin, type IV collagen, and fibronectin in normal and cryptorchid human testes. An immunohistochemical study
Authors: SANTAMARIA L. , MARTINEZ‐ONSURBE P. , PANIAGUA R. , NISTAL M.
Published: May 2008

An immunohistochemical study of laminin, type IV collagen, and fibronectin was carried out in the testes of normal men and in the cryptorchid and contralateral scrotal testes of cryptorchid men from 2 to 40 years of age. The integrated optical density (IOD) per unit area of the lamina propria was measured in the immuno‐stained sections. Fibronectin was found throughout the thickness of the lamina propria of the seminiferous tubules and in the interstitial connective tissue. No differences between normal and cryptorchid testes were found. Laminin was observed in the innermost part of the lamina propria of the seminiferous tubules and surrounding the endothelium of blood capillaries from infancy. No differences were found between normal and cryptorchid testes in the prepubertal period. In adult cryptorchid testes, laminin formed more numerous and deeper invaginations towards the seminiferous epithelium than in normal adult testes. Type IV collagen appeared throughout the thickness of the lamina propria of normal testes as well as in the wall of interstitial blood vessels. From infancy, the lamina propria of seminiferous tubules, but not blood vessel walls, showed lesser immunostaining for type IV collagen and a lower IOD of this component than did control tests from men of the same age. No differences between unilateral and bilateral cryptorchid‐ism were found. The contralateral scrotal testes of cryptorchid males showed intermediate immunostaining for type IV collagen between that of normal control testes and that of cryptorchid testes. These findings suggest that the lamina propria of seminiferous tubules is lesioned at an early age in both cryptorchid and contralateral scrotal testes of cryptorchid men.
Cloning and analysis of mRNAs expressed specifically in the human epididymis
Authors: KIRCHLOFF C. , OSTERHOFF C. , HABBEN I. , IVELL R.
Published: May 2008

Differential cDNA screening identified four new genes that are expressed specifically in the human epididymis. Their corresponding mRNAs are relatively short, ranging from 0.7 to 1.0 kb, and belong to a class of abundant or moderately abundant transcripts. Tissue‐specific expression was pronounced for all four sequences, which showed no hybridization signals with RNA from eight different human tissues. Cross‐hybridization with epididymal RNAs from some non‐rodent mammalian species was observed with two cDNAs; the remaining two transcripts failed to cross‐hybridize with any of the animal epididymal RNAs tested.
Age‐related differences in testicular microcirculation
Authors: DAMBER J. ‐E. , BERGH A. , WIDMARKt A.
Published: May 2008

Different aspects of testicular microcirculation were studied in rats aged 20, 24 or 90 days, in order to determine if maturational changes in the testis could influence its vasculature. Using laser Doppler flowmetry, it was found that vasomotion was not present in 20‐day‐old rats, but it could be induced by hCG‐treatment given 32 h prior to flow measurements. In rats aged 24 days vasomotion was present in the testis but it was not influenced by hCG treatment, in contrast to adult rats in which testicular vasomotion disappeared after hCG treatment. In all age‐groups, hCG treatment induced an increase in testicular interstitial fluid (IF) volume, which was most pronounced in young rats. In young rats, as in adults, there was an increase in the volume density of polymorphonuclear (PMN) leucocytes in testicular blood vessels after hCG treatment. However, rats aged 20 or 24 days did not exhibit any significant extravascular migration of leucocytes as occurred in adult rat testis after hCG injection. The deposition of colloidal carbon particles observed in adult rats after hCG‐treatment was also not seen in young rats. It is concluded that the vasculature of the immature rat testis reponds to hCG stimulation with an increase in IF volume. However, when compared to adult rats, different mechanisms may be operating because no evidence for opening of inter‐endothelial cell gaps was observed in the testis of young rats.
Anti‐spermatogenic activity of malvidin chloride in langur monkeys (Presbytis entellus entellus Dufresne)
Authors: BHARGAVA S. K.
Published: May 2008

Malvidin chloride (MC) a colouring agent from flowers of Malvaviscus conzattii Greenum was studied for male anti‐fertility effects in adult langur monkeys (Presbytis entellus entellus Dufresne). When fed 50 mg MC kg ‐1 for a period of 60 days, inhibition of spermatogenesis resulted. The weights of testes and epididymides were reduced and there was atrophy of the Leydig cells. In the epididymis, epithelial cell heights were reduced after MC‐treatment. Conspicuous shrinkage of seminiferous tubules and Leydig cell nuclei were evident. Depletion of total proteins, RNA, sialic acid, alkaline/acid phosphatase activity in testes and epididymides with the elevation of testicular levels of cholesterol and glycogen also occurred. Blood/serum analysis and haematology of MC‐treated langur monkeys revealed that all of the clinically important parameters were well within the normal range. The slightly increased bilirubin concentration and alkaline phosphatase activity returned to normal range within 30 days of the last dose of MC. The anti‐spermatogenic activity of MC in langur monkeys is discussed.
The levels and possible involvement of leukotriene B4 and prostaglandin F2α in the control of interstitial fluid volume in the rat testis
Authors: ABAYASEKARA D. R. E. , KURLAK L. O. , JEREMY J. Y. , DANDONA P. , SHARPE R. M. , COOKE B. A.
Published: May 2008

The possible involvement of two arachidonic acid metabolites, prostaglandin F2α (PGF2α) and leukotriene B4 (LTB4), in the stimulatory effect of human chorioni gonadotrophin (hCG) on the volume of interstitial fluid (IF) in the rat testis ha been investigated. Administration of hCG caused a time‐ and dose‐dependen increase in the IF levels of PGF2α while LTB4 showed no clear dose‐dependence but did decrease significantly at 2–8 h after injection of 100 IU hCG. Administra tion of ethane dimethane sulphonate (EDS), which specifically destroys Leydi cells, decreased the volume of IF but the IF levels of LTB4 and PGF2α were unchanged. This indicates that although the absence of Leydig cells results i significant changes in IF volume, LTB4 and PGF2α are probably not involved in these changes. Furthermore, these findings suggest that Leydig cells are not th only contributors to LTB4 and prostaglandins E2 and F2α in testicular IF. In rats injected peripherally with hCG, intratesticular administration of the cyclo oxygenase inhibitor, indomethacin, decreased PGF2α, levels in IF after 2 and 4 ’ but not at later times. However, no inhibitory effect of indomethacin on the hCG stimulated increase in IF volume was detected; indeed, at 2 h after injection o indomethacin + hCG there was a significant increase in IF volume in the indomethacin‐injected testis. It is concluded that, although prostaglandins am LTB4 are secreted into IF in the rat testis, these arachidonic acid metabolites ar apparently not the primary mediators of the increase in IF volume that follows hCC treatment.
Analysis of the relationship between reactive oxygen species production and leucocyte infiltration in fractions of human semen separated on Percoll gradients
Authors: AITKEN R. J. , WEST K. M.
Published: May 2008

We have shown previously that the reactive oxygen species generated by washed human ejaculates originate from cells which can be isolated in the low density region of discontinuous Percoll gradients. In this study we have used a simplified two‐step (40/80%) Percoll gradient to separate human ejaculates (n=109) into two populations of spermatozoa, exhibiting either a high or a low capacity for reactive oxygen species generation. We have then examined the relationships between this activity and other properties of the isolated fractions, with particular emphasis on the presence of leucocytes, which we have quantified using a monoclonal antibody directed against the common leucocyte antigen. The low‐density cells recovered from the 40%/80% interface of the Percoll gradients, differed from the high‐density fraction in exhibiting significantly reduced sperm motility, poorer sperm morphology, and a considerably enhanced capacity for reactive oxygen species production (P<0.001). In six cases the elevated levels of reactive oxygen species generation were associated with leucocyte concentrations in excess of 1 times 106 per 107 sperm, suggesting that leucocytes enter the seminal compartment in an activated, oxygen‐radical generating, state. However, in the majority of cases exhibiting high levels of reactive oxygen species production, leucocyte numbers were low or absent and the semen profiles were unremarkable, except that seminal sperm concentrations tended to be low. These results suggest that the oxidative stress responsible for defective sperm function involves reactive oxygen species originating from two sources; the sperm and infiltrating leucocytes.
Aniline blue staining as a marker of sperm chromatin defects associated with different semen characteristics discriminates between proven fertile and suspected infertile men
Authors: AUGER J. , MESBAH M. , HUBER C. , DADOUNE J. P.
Published: May 2008

A retrospective study of 49 men with proven fertility and 396 suspected infertile men was conducted with the primary objective of investigating the relationship between the nuclear maturity of sperm and male fertility. Acidic aniline blue staining was used to detect chromatin defects of sperm nuclei related to their nucleoprotein content as associated with DNA. The discriminant value of the percentage of unstained nuclei (= percentage of mature heads, MH) and of other semen characteristics, was analysed by a stepwise (forward) linear regression model. Semen characteristics that discriminated significantly between the two groups of subjects were, in descending order: (1) the percentage of normal sperm, (2) the percentage of amorphous heads, (3) the percentage of tapered heads, (4) semen volume, and (5) the percentage of MH. The discriminant value of each of the significant characteristics was studied by means of ROC‐curves. MH had the best ROC‐curve profile; its cut‐off value was found to be equal to 70% (74.5 ± 2.6% for the donor group versus 53.0 ± 1.1% for the patient group). A simple infertility score (SIS) including MH, was built according to the cut‐off values inferred from the ROC‐curves. SIS allowed an overall satisfactory separation of the two groups (≤4 = fertile, 5–6 = uncertainty zone, >6 = infertile). Our results indicate that the addition of the evaluation of sperm head maturity to routine semen analysis improves the assessment of fertility in men.
Carcinoma‐in‐situ cells in cultured human seminiferous tubules
Authors: LAUKE H. , SEIDL K. , HARTMANN M. , HOLSTEIN A. F.
Published: May 2008

For the first time, early germ‐cell derived tumour cells were studied in an in‐vitro system of cultured seminiferous tubules. The intratubular tumour cells not only survived in culture for 7 days but were also able to multiply. Dividing tumour cells were identified in semi‐thin sections and electron micrographs by morphological criteria. Additionally, mitotic activity was demonstrated by ‘Hlthymidine histo‐autoradiography’. There are numerous reports on cell lines established from solid non‐seminomas, but up to now no references to seminoma cell lines or cultures of intratubular tumour cells are available. The culture of seminiferous tubules offers a tool in making carcinoma‐in‐situ cells accessible for experimental work.
Effects of seminal plasma from normal and asthenozoospermic men on the progressive motility of washed human sperm
Authors: MAGNUS Ø. , BREKKE I. , ÅBYHOLM T. , PURVIS K.
Published: May 2008

Human seminal plasma stimulated the progressive motility of human sperm in a dose‐ and time‐related manner. Serum exhibited a similar capacity for stimulation. Seminal plasma from different normozoospermic men showed marked variation in its capacity to stimulate sperm motility. The stimulatory effect was maintained after heat‐treatment, indicating the action of a low molecular weight substance or metal ion. No differences could be observed in the capacity for stimulation between seminal plasma from normozoospermic (n = 23) and asthenozoospermic (n = 22) men, when tested at the same dilution and under identical conditions. It is concluded that in general, differences in seminal plasma composition cannot account for the reduced sperm motility in asthenozoospermic men. Furthermore, the stimulatory effects of seminal plasma may be a property shared by other biological fluids.
Variation of semen quality in normal men
Authors: MALLIDIS C. , HOWARD E. J. , BAKER H. W. G.
Published: May 2008

To examine the amount of variability in semen analysis results and whether there is any effect of season, 673 specimens provided by seven normal, healthy men (61–205 specimens/subject) over 72–324 weeks were assessed for sperm concentration, ejaculate volume, motility and motility index. Noticeable sample to sample variations were found. The largest proportion of the overall variance was due to within‐subject differences, e.g., sperm concentration (54%), ejaculate volume (59%), percentage motility (96%) and motility index (74%). Although changes in semen‐analysis results occurred over the year, no consistent trend was seen. No evidence was found to suggest that the differences were due to modifications of the methods employed by the laboratory, or to the change of season.
The effect of A23187 concentration and exposure time on the outcome of the hamster egg penetration test
Authors: FORD W. C. L. , REES J. M. , MCLAUGHLIN E. A. , GODDARD R. J. , HULL M. G. R.
Published: May 2008

We investigated the conditions required to enhance the performance of human sperm in the hamster egg penetration test with the free acid form of A23187. The best performance was observed after stimulation with 2 μM A23187 for 1 h when the median penetration rate with sperm from fertile donors was 100% of eggs with 5.8 decondensed sperm headdegg. Extending the stimulation period with 2 μm A23187 to 2 or 3 h, resulted in a progressive decrease in the penetration rate. In the absence of A23187, the penetration rate was lower (0.7 decondensed sperm heads/ egg after 1 h) but increased with stimulation time. A similar picture was observed with sperm from patients taken for an IVF programme. For a pool of cryopreserved semen, the coefficient of variation of the penetration rate after stimulation with 2 μM A23187 for 1 h, expressed as decondensed sperm headdegg, was 11% within and 20% between assays. There was no correlation between the outcome of the hamster egg penetration test and the percentage motility, velocity or lateral head displacement of the sperm measured after the same stimulation regime. However, in IVF patients the initial velocity and lateral head displacement of the sperm (zero time) were correlated with the best result from the hamster egg penetration test (r= 0.62 and 0.57 respectively). No motility changes characteristic of capacitation were detected. We conclude that stimulation with 2 μM A23187 (free acid) for 1 h prior to the addition of the zona free hamster eggs can produce a high penetration rate with fertile samples and provides a convenient and robust protocol for the assay. However, when carried out in this way the test does not assess the ability of the sperm to capacitate.
Detection of antisperm‐antibodies in patients with primary testicular cancer
Authors: FOSTER R. S. , RUBIN L. R. , MCNULTY A. , BIHRLE R. , DONOHUE J. P.
Published: May 2008

Serum antisperm‐antibodies have been noted to develop as a result of various types of testicular pathology and injury. Their presence sometimes correlates with impaired fertility. We examined 52 patients with non‐seminomatous low‐stage testicular cancer for the presence of serum antisperm‐antibodies to determine whether or not auto‐immunity is a factor in the depressed fertility known to exist in these patients. Twenty‐one per cent were discovered to have serum antisperm‐antibodies using an immunofluorescent technique.
Selective action of androgens on the molecular forms of esterases characterized by two‐dimensional gel electrophoresis in the epididymis and vas deferens of the mouse
Authors: ABOU‐HAÏLA AÏDA , MARCELLE‐ANNE  , FAIN‐MAUREL
Published: May 2008

Molecular forms of esterases were resolved in non‐denaturing conditions by using two‐dimensional gel electrophoresis with isoelectric focusing in the first dimension and a time‐dependent polyacrylamide gradient gel electrophoresis (PAGGE) in the second dimension. This procedure was used to analyse sequential changes in esterase composition along the excurrent genital duct of the mouse and to initiate a specific identification of the androgen‐regulated molecular forms. Almost all the 68 variants (pH 3.9–6.4 and 50–300 kDa) revealed by α‐naphtyl acetate from the fluids of the three parts of the epididymis (caput, corpus, cauda) and vas deferens, could be assigned to the carboxylesterase group as shown by their action on various substrates and sensitivity to inhibitors. Some of these variants co‐migrated with those in the serum and testis, whereas other enzyme forms made their first appearance in the caput (13), in the corpus (26) and in the vas deferens (3). The major changes occurred between the caput and the corpus of the epididymis. Only a few acidic spots were not revealed after neuraminidase digestion. Castration of mice (4 weeks) resulted in inhibition of the activity of 34 esterase forms, and thus abolished most of the regional differences in the excurrent duct system. By re‐initiating or repressing the synthesis of regional esterase variants, testosterone supplementation (2 and/or 4 weeks) of castrated animals restored the normal esterase pattern in the three epididymal parts, but not in the vas deferens. The major effect of efferent duct ligation (4 weeks) was the emergence in the corpus and cauda of the epididymis of two variants found in the caput of uncastrated mice.
Production and secretion of an interleukin‐l‐like factor is stage‐dependent and correlates with spermatogonial DNA synthesis in the rat seminiferous epithelium
Authors: SÖDER O. , SYED V. , CALLARD G. V. , TOPPARI J. , PÖLLÄNEN P. , PARVINEN M. , FRÖYSA B. , RITZÉN E. M.
Published: May 2008

It has been shown previously that the intact adult rat testis produces large amounts of an interleukin‐1 (IL‐1)‐like growth factor. The present study has investigated whether this testicular IL‐1‐like factor (tIL‐1) is produced and secreted differentially by the fourteen stages of the seminiferous epithelial cycle in the rat testis. Seminiferous tubule segments representing defined stages were identified by tran‐sillumination‐assisted microscopy and isolated by microdissection. Pooled segments were either homogenized and extracted with aqueous buffer or incubated for 24 h to produce conditioned media (CM). The recovered material was then analysed for IL‐1 bioactivity in a sensitive murine thymocyte proliferation assay. When divided into four stage groups, extracts of stages II‐VI, IX‐XII and XIII‐I showed equally high IL‐1 activity whereas stage group VII‐VIII showed much lower activity. More detailed analysis with 10 different stage groups showed that tIL‐1 activity was undetectable in extracts of substages VIIab and VIIcd. The same pattern was seen when CM from cultured tubular segments were analysed. Labelling of seminiferous tubules with tritiated thymidine in vitro and analysis by auto‐radiography revealed that DNA‐synthesizing spermatogonia were absent in sub‐stages VIIb and VIIc and sparse in substages VIIa and VIId. The results show that tIL‐1 activity is produced in a stage‐dependent manner and suggest that tIL‐1 might be involved in the regulation of spermatogonial proliferation in vivo.
The fertility potential of male cystic fibrosis patients
Authors: Gottlieb C. , Plöen L. , Kvist U. , Strandvik B.
Published: May 2008

Men suffering from cystic fibrosis (CF) are considered to be infertile because of azoospermia. In testicular biopsies from two patients with CF normal spermatogenesis was found despite the absence of sperm in the ejaculate. Fructose and prostaglandin were not detectable in the semen whereas the levels of acid phosphatase and zinc were within normal limits, indicating normal prostatic function and absence of seminal vesicles. These findings may improve the possibility for male patients with CF to father a child.
Testicular lesions and lymphocyte subpopulations in rats immunized with a soluble fraction of testicular homogenate
Authors: Doncel G.F. , Denduchis B. , Lustig L.
Published: May 2008

A soluble fraction obtained from a testicular homogenate by precipitation with ammonium sulphate (ASPM) was emulsified with Freund's complete adjuvant (CFA) and injected into Wistar rats. At 50 days after the first immunization (total of three injections) the animals had developed moderate and multifocal testicular damage, characterized mainly by sloughing of the seminiferous epithelium. A delayed‐type hypersensitivity response and circulating antibodies to ASPM were detected at different times with maximum levels at 50 days. The addition of Bor‐detella pertussis to the immunization did not increase the severity of the lesion but augmented the cellular and humoral immune response to ASPM. The phenotypic characterization of cells present in the lymph nodes draining from the site of immunization in animals injected with CFA alone (control group) revealed an increase in CD8+ T‐cells and a low CD4/CD8 ratio. Conversely, rats immunized with CFA plus ASPM (experimental group) exhibited testicular damage and showed a significant decrease in CD8+ cells with a normal CD4/CD8 ratio. In conclusion, rats immunized with a testicular antigen developed focal aspermatogenic lesions and a concomitant specific immune response as well as lymph‐node cell variations focused apparently on the CD8+ T‐cell subpopulation.
Pattern of Sertoli cell degeneration in cryptorchid prepubertal tests
Authors: RUNE G. M. , MAYR J. , NEUGEBAUER H. , ANDERS CH. , SAUER H.
Published: May 2008

Seventy‐three testicular biopsies from 54 children (aged 2 months‐14 years) with undescended testes were examined by light and electron microscopy. The biopsies included abdominal, inguinally fixed, inguinally moveable, and retractile testesl. Alterations in Sertoli cell morphology were found in all biopsies. The alterations included dilated elements of rough endoplasmic reticulum, vacuolization of the cytoplasm, mitochondria with poorly preserved cristae, increase in electron density of the matrix, elongation of the nuclei, and irregularities of the nuclear membrance. According to the numerical appearance of these cells and to the extent of lesions in single Sertoli cells, seven phases in the contionuous process of tubular alteration were distinguished. The most severe tubular damaged (phase VII> occurred when the seminiferous epithelium consisted exclusively of necrotic cells. All phases of tubular alterations were seen regularly in each of the biopsies investigated. Germ cells occurred only in phases I‐IV and were never observed in tubules in phases V‐VII. Significant differences became evident between inguinal and retractile tests by morphometric evaluation. It was demonstrated that the number of germcells per cross‐sectioned tubule (S/T value) correlated negatively with the percentage of tubules in phases V‐VII. In contrast to inguinal testes, a complete absence of Sertoli cells and an S/T value <0.1 were never found in retractile tests and the percentage of tubules in phases V‐VII was reduced signficantly compared with inguinal testes.

Our findings indicate that (i) maldescended testis in patients between 1 and 15 years‐of‐age is associated with a special pattern of Sertoli cell degeneratlion; (ii) Sertoli cell degeneration is a continuous process, which can lead eventually to complete dissolution of the seminiferous epithelium; (ii) total degeneration is not related to age but is dependent on testicular pposition; (iv) a defined phase of degeneration excludes germ cell development, and therefore enhanced Sertoli cell degeneration in cryptorchid tests must also account for the reduction in germ cell number.
The role of the seminal vesicles and coagulating glands in fertilization in the rat
Authors: SOFIKITIS N. , TAKAHASHI C. , KADOWAKI H. , SHIMAMOTO T. , NAKAMURA I. , MIYAGAWA I.
Published: May 2008

To investigate the role of the seminal vesicles (SV) and coagulating glands (CG) in fertilization in the rat, partial resections of the SV (25|X%–group B), 50|X%–group E, and 75|X%–group D) or bilateral resection of the SV alone (100%|X%group E), or together with the CG (group F) were undertaken. In other groups, bilateral resection of the CG only (group G), or bilateral resection of the CG with ipsilateral SV resection (group H) were performed. A group of sham‐operated rats served as controls (group A). There were no significant differences among groups A, B, C and D in the number of fertile rats post‐operatively but significant differences were evident in groups E, F, G, H when compared with the control group (P<0.01 for all groups). Only the fertile rats ere able to form copulatory plugs (CP) post‐operatively.

Bilateral ligation of the SV ducts was performed in ten fertile rats. Post‐operatively, the proportion of fertile rats was decreased significantly (P<0.01). Removal of the ligatures resulted in a significant increase (P<0.01) in fertility. It was also demonstrated that the addition of rat seminal vesicular secretions (SVS) to epididymal sperm suspensions resulted in a significant decrease (P<0.001) in sperm motility.

These results suggest that the SV and CG are necessary for fertility in rats, and that one role of SVS is the formation of a CP and not the maintenance of sperm motility.
Sperm chromatin stability and zinc binding properties in semen from men in barren unions
Authors: KJELLBERG S. , BJÖRNDAHL L. , KVISTt U.
Published: May 2008

Sperm chromatin stability and zinc binding properties were studied in semen samples from 115 men living in barren unions. Of these men, 26|X% had a high proportion of swelling sperm, i.e. less than 80|X% sperm with stable chromatin after exposure to the detergent sodium dodecyl sulphate. From 2–67|X% of seminal zinc was bound to high molecular weight ligands of vesicular origin (HMW). This shows that, among infertile men, liquefied seminal plasma has huge variations in zinc chelating properties. The relationship between prostatic palpatory status, the proportion of abnormal sperm, the percentage zinc bound to HMW (HMW‐Zn), the time between ejaculation and analysis and chromatin stability were studied. Samples with low chromatin stability were found more frequently in men with low HMW‐Zn levels in semen. The proportion of stable sperm decreased in samples with prolonged exposure to seminal plasma. Neither the proportion of stable sperm heads nor the percentage zinc bound to HMW could be used to predict the future chances of the infertile men fathering children when studied 15–180 min after ejaculation. To differentiate between initial zinc‐dependent stability and superstability developed in seminal plasma, other more sensitive methods must be developed.
Management of the non‐descended testis: doubtful value of luteinizing‐hormone‐releasing‐hormone (LHRH). A double‐blind, placebo‐controlled multicentre study
Authors: OLSEN L. H. , GENSTER H. G. , MOSEGAARD A. , JØRGENSEN F. S. , HOFMAN N. , JENSEN V. B. , LASSEN L. B. , RASSMUSSEN M. , VINZENTS L. , DAMMEGAARD L. , MUNCK S. , BRÖNDUM‐NIELSEN K.
Published: May 2008

In a double‐blind, placebo‐controlled multicentre study, the effect of luteinizing‐hormone‐releasing‐hormone (LHRH) in 141 boys was analysed after 4–week treatment period with 0.4 mg LHRH nasal spray or placebo nasal spray three times daily. Data from 123 boys was analysed, with 62 boys in the treatment group and 61 in the placebo group. Full response i.e. the testis at the bottom of the scrotum on both sides in boys with bilaterally undescended testes, was found in six patients, one of them in the placebo group [Therapeutic gain of LHRH with 95% CI: 8.1% (0.1–16.6%, P = 0.12)]. Only in these boys could surgery be avoided. Considering the number of testes (and not the number of boys) a significant effect was found on at least one testis in 25% of boys with bilaterally undescended testes [Therapeutic gain with 95% CI: 24.0% (13.2–34.8%, P = 0.001)]. In unilateral undescended testes, the LHRH treatment showed no effect (P = 1.00). The inclusion of retractile testes did not affect our results. In our opinion LHRH has a limited place in treatment of the non‐descended testis.
Effect of different culture techniques used to induce capacitation on the chromatin stability of human sperm
Authors: OLIVA A. , PORSTMANN E. , CAILLE A. , BARTOLOMEO C.
Published: May 2008

To validate an in‐vitro bioassay for assessing chromatin stability of human sperm, 38 semen samples from infertile men were studied using sodium dodecyl sulphate, an anionic detergent which disorganizes only the cytoplasmic membrane. Assay sensitivity was 50 sperm, whilst the within‐ and between‐assay variation, and the between‐observer variation were found to be within the accepted range for this type of bioassay. The influence of different in‐vitro treatments currently used in some clinical assisted fertilization programmes was evaluated: a destabilizing effect occurred in Grade I (stable) and Grade III (swollen) sperm. In the former, all treatments reduced stable sperm; in the latter, a significant (P<0.001) increase in swollen sperm was shown with two methods that used Ham's F‐10 as culture medium. Different chromatin patterns found in the treated sperm suggest the possibility that the recovered samples could be modified compared to their status at the time of isolation.
Therapy with human chorionic gonadotrophin alone induces spermatogenesis in men with isolated hypogonadotrophic hypogonadism‐long‐term follow‐up
Authors: VICARI E. , MONGIOÌ A. , CALOGERO A. E. , MONCADA M. L. , SIDOTI G. , POLOSA P. , D'AGATA R.
Published: May 2008

The effects of long‐term (14–120 months) hCG‐treatment of 17 male patients affected by isolated hypogonadotrophic hypogonadism (IHH) on testicular volume, plasma testosterone levels, and sperm concentration were assessed. Mean testicular volume increased from 3.8 ± 0.2 (Mean ± SEM) ml to a maximal of 14.9 ± 1.1 ml after 22.2 ± 2.3 months of hCG treatment. Maximal testicular volume correlated positively with the volume recorded before the patients had undergone any previous treatment. Testicular growth was also analysed by sorting the patients into two sub‐groups according to whether their initial testicular volume was <4 ml (small testis subset, STS) or ≥4 ml (large testis subset, LTS), supposedly indicating complete or partial gonadotrophin deficiency, respectively. Testicular volumes in the LTS group were always greater than those of the STS. Plasma testosterone levels reached adulthood values during hCG treatment and no statistically significant difference was detected between LTS and STS patients with IHH. Thirteen patients (70%) became sperm‐positive during treatment with hCG alone; five out of eight (60%) were STS patients and eight out of nine (90%) were LTS. In addition, LTS patients always had a greater sperm output than did STS patients. Sperm concentration correlated positively with maximal testicular volume, but not with patient age, length of treatment, or initial testicular volume. The administration of hMG to eight of these patients caused an increase in testicular volume in two patients but the mean volume was not statistically different from that recorded at the end of treatment with hCG alone. Similarly, sperm concentration improved in three patients but again it did not differ significantly from that achieved in the course of hCG treatment. It is noteworthy that one patient became sperm‐positive after the addition of hMG to his therapeutic regimen. Among sperm‐positive patients attempting conception, seven out of 10 succeeded, two of whom were from the STS group.

In summary, this study indicates that hCG alone is an effective treatment to induce complete spermiogenesis in IHH patients regardless of their initial testicular volume. However, a number of IHH patients may benefit from the addition of hMG in terms of testicular volume, sperm output, and pregnancy outcome.
Efficacy of varicocele embolization versus ligation of the left internal spermatic vein for improvement of sperm quality
Authors: YAVETZ H. , LEVY R. , PAPO J. , YOGEV L. , PAZ G. , JAFFA A. J. , HOMONNAI Z. T.
Published: May 2008

Efficacy of surgical varicocelectomy versus embolization of the spermatic vein was studied in 137 men diagnosed as suffering from left varicocele. The men were divided randomly into three groups according to the methods of treatment: A ‐embolization of the internal spermatic vein (51 men); B ‐ Ivanissevich technique of high ligation of the spermatic veins (43 men); and C ‐ Bernardi technique of high ligation (43 men). The groups were similar in terms of age, duration of infertility and possessed semen characterized as oligoteratoasthenozoospermia. The fertility of the female partners was evaluated carefully and they were found to be potentially fertile. Varicocele was diagnosed by at least two of the following methods: physical palpation during valsalva manoeuvre, venography, or scrotal scanning using the technetium pertechnate radioactive method. Semen quality was assessed before treatment and at 3, 6 and 9 months post‐treatment. Fecundity was followed‐up for 18 months. The major results were:

Shrinkage of the varicocele was found in all three groups studied. The same rate of recurrence was recorded in the three groups (24%, 37% and 35% in groups A, B and C, respectively).

Improvement of sperm quality was significant in groups A and B, with better results in group B.

The pregnancy rate was significantly higher in group B, compared with A (38.2% vs. 20.6%; P <0.05).

Thus, high ligation of the internal spermatic vein yields better results than low ligation or embolization as far as semen quality and pregnancy is concerned.
Temporal changes in the serum levels of gonadotrophins and testosterone in male rats bearing subcutaneous implants of 5α‐dihydrotestosterone
Authors: PARTE P. , JUNEJA H. S.
Published: May 2008

This study has assessed the effect of s.c. implants of 5a‐dihydrotestosterone (DHT) on the blood levels of testosterone and gonadotrophins in intact and castrated adult male rats. The rats were bled via cardiac puncture at 1, 3, 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 days after DHT implantation. On days 28 and 49 post‐implantation, rats were injected with LHRH (25 ng) and bled 15 min later. In intact rats bearing DHT implants, the serum levels of LH and testosterone were suppressed significantly with no significant changes in FSH levels. Ventral prostate, seminal vesicles and the pituitary were reduced significantly in weight when compared with controls with empty implants. DHT significantly inhibited LHRH‐induced release of FSH in intact rats. In castrated rats, DHT implants inhibited the secretion of both LH and FSH, with a rise in serum DHT levels. DHT stimulated the LHRH‐induced release of LH but inhibited FSH. DHT implants increased the weight of the seminal vesicles and ventral prostate but inhibited the weight of the pituitary when compared to castrated rats bearing empty implants. This study demonstrates specific inhibition of serum LH and testosterone by DHT implants in intact adult rats.
Does follicle‐stimulating hormone or pregnant mare serum gonadotrophin influence testicular blood flow in rats?
Authors: BERGH A. , DAMBER J‐E. , LIEU L. , WIDMARK A.
Published: May 2008

Treatment of adult rats with 25 iu follicle‐stimulating hormone (FSH) did not influence testicular blood flow, serum testosterone, vasomotion or intravascular leucocyte concentration at 6, 12 and 24 h after treatment. Treatment of adult rats with 50 iu pregnant mare serum gonadotrophin (PMSG) resulted in a two‐fold increase in testicular blood flow at 24 h, and an increase in interstitial fluid volume at 36 h after treatment. This PMSG‐induced increase in blood flow did not occur in Leydig cell‐depleted animals, suggesting that the effect on blood flow is mediated via the Leydig cells. PMSG injection stimulated testosterone secretion but it did not influence vasomotion, and it only marginally increased the secretion of leuco‐tactic factors in the testis. The present study suggests that FSH has no apparent effects on testicular blood flow and that the effects of PMSG (a hormone with both FSH and LH‐like activity) is mediated via stimulation of the Leydig cells.