bluuebunny/crossref_metadata_2025 huggingface dataset:
Testosterone concentrations in human seminal plasma and saliva and its correlation with non‐protein‐bound and total testosterone levels in serum
Authors: Sannikka E. , Terho P. , Suominen J. , Santti R.
Published: May 2008

A sensitive, specific and precise non‐chromatographic method for the radio‐immunoassay of testosterone in human seminal plasma and saliva from adult and pubertal males is described, and the values compared to total and non‐protein‐bound testosterone levels in serum. There was a significant correlation between salivary and serum‐free levels of testosterone (r = 0.75, P < 0.001, n = 67) whilst the correlation of serum levels of total testosterone with free as well as with salivary testosterone levels was weaker (r = 0.63 and 0.64, respectively). The salivary and serum levels of free testosterone showed better correlation with the stage of puberty than did the serum levels of total testosterone. Further evidence for a correlation between salivary and serum levels of free testosterone was obtained following oral administration of testosterone undecanoate, as this treatment increased the mean concentration of serum total testosterone after 3 h by 82%, but increased salivary and serum levels of free testosterone by only 30% and 20%, respectively. The coefficient of correlation between serum levels of total testosterone and seminal plasma testosterone was 0.73 (P < 0.001), whilst the correlation between levels of serum‐free testosterone with both salivary and seminal plasma levels of testosterone was statistically non‐significant. Our observations on salivary testosterone are in accordance with the diffusion of non‐protein‐bound steroids into peripheral tissues, and consequently into their secretions. This model, however, does not appear to be applicable to the sex accessory glands.
Microbiological investigations on cryopreserved human semen
Authors: Glander Hans‐Jürgen , Schönborn Christina , Rytter Manfred
Published: May 2008

Primary mycological contamination was studied in 565 random human ejaculates. After sperm preservation at low temperature, secondary mycological contamination of 55 of the ejaculates was investigated. 25 ejaculates were cryopreserved with and without antibiotics and examined bacteriologically before and after different phases of cryopreservation. Furthermore, semen samples of several donors were inoculated with suspensions of different concentrations of yeasts and with the test bacteria Escherichia coli K 12 and Staphylococcus aureus SG 511. Microbiological results were then compared before and after cryopreservation. The following results were obtained: 1. 9.8% of samples showed primary mycological contamination (1.9% with Candida albicans). 2. Cryopreservation reduced the concentration of fungi by more than 90% on average. 3. Only one semen sample was contaminated with bacteria (Staphylococcus aureus haemolyticus and Streptococcus nonhaemolyticus). Non‐pathogenic bacteria were found in 11 ejaculates (Micrococcus luteus in 5 samples and Staphylococcus epidermidis in 6 samples), and contamination was largely unaffected by cryopreservation, even in the presence of antibiotics. 4. Test bacteria inoculated into semen were not influenced by the addition of antibiotics to the cryoprotective medium.
Selective uptake by the Sertoli cells of cytoplasm from normal spermatogonia in the rat testis
Authors: Ulvik Nils Magnus
Published: May 2008

Electron microscopical examination of germ cells during their development from early type A spermatogonia to late pachytene spermatocytes showed that small, spherical pseudopodia emerged from type B spermatogonia and, to a lesser degree, from intermediate spermatogonia and early spermatocytes. Serial sections showed that the pseudopodia pinched off from the type B spermatogonia and were engulfed by the adjacent Sertoli cells. Groups of dense bodies were found in the Sertoli cells adjacent to the engulfed islands of germ cell cytoplasm. At a few instances islands of germ cell cytoplasm were seen to fuse with dense bodies in the Sertoli cells. The fate of the cytoplasmic islands is unknown, but phagocytosis by the Sertoli cells may be suggested. The findings indicate a new type of interaction between Sertoli cells and certain classes of spermatogonia.
Changes from puberty to adulthood in the concentration, motility and morphology of mouse epididymal spermatozoa
Authors: Albert Martine , Roussel Claude
Published: May 2008

A systematic study of the concentration, motility and morphology of epididymal spermatozoa was undertaken in mice of the OF1 strain, in order to characterize the changes observed during puberty. The comparative development of these 3 parameters was followed from days 30 to 90. A detailed morphological system of classification was established covering both individual and multiple abnormalities. At puberty, the first spermatozoa to appear were few in number, showed poor motility and were extremely atypical. Subsequently, the number of atypically‐shaped spermatozoa diminished, and their concentration and motility exhibited parallel increases. At 60 days, the values for these 3 parameters and the proportions of normal and abnormal spermatozoa became stable. During puberty, various forms of disruption of the midpiece as well as the presence of extremely atypical detached flagella resulted in a special pattern of sperm morphology, although all of these abnormal features disappeared at 40 days.
Prolactin secretion in the human male is increased by endogenous oestrogens and decreased by exogenous/endogenous androgens
Authors: Gooren L. J. G. , van der Veen E. A. , van Kessel H. , Harmsen‐Louman W. , Wiegel A. R.
Published: May 2008

There is evidence that prolactin may be involved in testicular steroidogenesis, and we have therefore investigated whether there is feedback regulation of androgens/oestrogens on prolactin secretion in the human male. To assess this we have measured basal and TRH‐stimulated prolactin levels in:

A) Six eugonadal men before and after 2 weeks' administration of the aromatase inhibitor Δ'‐testolactone, which led to a fall in oestradiol levels with unchanged levels of testosterone. In these patients, prolactin levels decreased.

B) Six eugonadal subjects before and after 2 weeks' administration of dihydrotestosterone undecanoate. In these subjects, prolactin levels decreased.

C) Six agonadal subjects, tested after 12 weeks' treatment with dihydrotestosterone undecanoate and compared to

D) Six agonadal subjects who received no sex steroid treatment. Again, it was found that dihydrotestosterone treatment decreased prolactin levels in patients from Group C.

E) Six eugonadal subjects were also studied before and after 6 weeks' administration of the androgen receptor antagonist, spironolactone, and this treatment increased Prl secretion.

It is concluded that in the human male, endogenous oestrogens increase prolactin secretion whilst exogenous/endogenous androgens decrease prolactin secretion.
Foetal testicular steroidogenesis and responsiveness to LH in freemartins and their male co‐twins
Authors: Shore L. S. , Shemesh M. , Mileguir F.
Published: May 2008

Foetal gonads were obtained from 1) 8 male foetuses that were co‐twin to freemartins, 2) 8 isosexual twins, 3) 59 singletons of 45–75 days of gestations, 4) 5 isosexual and 5 male co‐twins of 90–120 days and 5) 5 freemartins at 70–120 days of gestation. The gonads were incubated in supplemented medium 199 for 24 h in the absence or presence or LH and the testosterone and progesterone produced measured by RIA. During the time of sexual differentiation (45–75 days) the testes of the male co‐twins produced significantly less testosterone than testes from isosexual or singletons of the same age. Testes of foetuses older than 90 days were refractory to LH‐stimulation, but freemartin ovaries and testes from their male co‐twins continued to respond to LH with increased testosterone production. No significant differences in progesterone production were detected amoung co‐twins, isosexual twins or singletons at any age. We conclude that the testes of the male twin that is co‐twin to a freemartin displays abnormal steroidogenesis. This may be related to reports of abnormal testes after birth in males co‐twin to free‐martins.
Seminal plasma characteristics during frequent ejaculation
Authors: Rui H. , Gerhardt P. , Mevåg B. , Thomassen Y. , Purvis K.
Published: May 2008

Seven young men provided a semen sample every 8 h for 2 days. Citric acid, zinc, acid phophatase, spermine, fructose and protein were measured in the samples. Despite a dramatic reduction in semen volume, the concentration of the majority of the parameters remained constant during the period of study. The protein profile of the ejaculates, as assessed by 2‐dimensional electrophoresis, also did not change. Levels of citric acid, zinc and acid phosphatese were closely correlated, while spermine, which is also believed to be of prostatic origin, showed no correlation with these other parameters. The spermine concentration of the ejaculates increased significantly during the period of high ejaculation frequency and returned to normal levels after a 3 day abstinence period. In view of the fact that spermine accumulation in tissues appears to be associated with growth and protein synthesis this increase in seminal spermine may reflect a stimulation of prostatic cell activity.
Maturation of the juvenile rat testis after surgical treatment of cryptorchidism
Authors: Karpe B. , Plöen L. , Ritzén E. M.
Published: May 2008

Rats were made bilaterally cryptorchid at 17 days of age and bilateral orchidopexy performed at 34 days of age. The epididymal content of androgen binding protein (ABP), the weight and morphology of the testis, the cross‐sectional area of seminiferous tubules and the testicular concentration of testosterone were then studied at 34, 42, 59 and 120 days of age. Cryptorchidism was followed rapidly by progressive inhibition of spermatogenesis and testicular growth as well as by decreased Sertoli cell secretion of ABP. Orchidopexy resulted in a gradual restoration of spermatogenesis, and all impaired parameters seemed to improve at the same, fairly slow rate. Restoration was not complete, but by 120 days of age the morphological appearance of the testis was compatible with recovery of normal fertility.
Comparative pharmacokinetics of testosterone enanthate and testosterone cyclohexanecarboxylate as assessed by serum and salivary testosterone levels in normal men
Authors: Schürmeyer Th. , Nieschlag E.
Published: May 2008

The pharmacokinetics of 2 testosterone esters, testosterone enanthate and testosterone cyclohexanecarboxylate, were compared in a single blind crossover study in healthy young men. Their effects on serum and salivary levels of testosterone, as well as on the serum levels of LH, FSH and proclactin were measured after the injection of doses equivalent to 140 mg free testosterone. Both preparations yielded supraphysiological testosterone levels in serum and saliva as early as 2 h following injection, reaching peak levels 4 to 5 times above basal between 8 and 24 h. LH and FSH levels were suppressed as long as serum testosterone levels were elevated. Nine days after injecting testosterone enanthate and 7 days after giving testosterone cyclohexanecarboxylate, serum and salivary levels of testosterone had returned to basal. The longer activity of testosterone enanthate was also evidenced from more extended suppression of gonadotrophin levels. Although neither preparation is ideal because of the initial supraphysiological peaks, testosterone enanthate appears preferable for clinical use because of its slightly longer duration of action.
Levels of seminal free L(‐) carnitine in fertile and infertile men
Authors: Soufir J. C. , Ducot B. , Marson J. , Jouannet P. , Feneux D. , Soumah A. , Spira A.
Published: May 2008

Data about the levels of free L‐carnitine, an epididymal marker in human semen, are contradictory and unclear, particuliarly in their releationship to fertility. This can perhaps be explained by the absence of any studies of seminal L‐carnitine in a large group of fertile men, and by the lack of consideration of factors influencing its secretion.

In this study, free L‐carnitine was determined using a spectrophotometric method in deproteinized semen samples from fertile (n = 162) and infertile men without azoospermia (n = 303).

Our results can be summarized as follows:

Infertile men were found to have significantly lower (P < 0.001) seminal carnitine levels (755 ± SD 499 nmoles) compared with fertile men (1010 ± 570).

Percentiles have been calculated for fertile men, and ‘normal’ values proposed (10th percentile = 390 and 90th percentile = 1830 nmoles).

Distribution of the levels of L‐carnitine were asymmetric in fertile as well as in infertile men (median: 922 nmoles vs 645).

In both groups, a significant increase in carnitine levels was observed with increasing length of abstinence, and a decrease in the ratio of carnitine/days of abstinence.

Methodological, physiological and pathological factors which may explain these results are discussed.
Stimulatory effect of an LHRH‐agonist on testicular blood flow in hypophysectomized rats
Authors: Damber J.‐E. , Bergh A. , Daehlin L.
Published: May 2008

The effect of an LHRH‐agonist (A) on testicular blood flow and plasma testosterone levels was studied in adult hypophysectomized rats. Subcutanous injection of 1 μg LHRH‐agonist 2 or 4 h prior to experiment induced an increase in testicular blood flow. (Controls: 21.1 ± 2.8 ml 100 g x min; LHRH‐A + 2h: 28.9 ± 3.3 ml/100 g x min; LHRH‐A + 4h: 32.4 ± 4.1 ml 100 g x min P < 0.05; all mean ± SEM). There was also a significant increase in plasma testosterone levels at 2 h (P < 0.01) and 4 h (P < 0.01) after treatment with the LHRH‐agonist, with a peak at 2 h.
The effect of an 1100 km run on testicular, adrenal and thyroid hormones
Authors: Schürmeyer T. , Jung K. , Nieschlag E.
Published: May 2008

Although endocrine effects of physical or psychological stress are well documented, it is not known to what extent adaptation to prolonged exertion occurs. We therefore investigated the impact of an 1100 km run of 20 days' duration on selected pituitary, testicular, adrenal and thyroid hormones. Blood samples were obtained from five male athletes prior to and after the day's run on the 1st, 5th, 9th, 14th and 19th day. Results show that adrenal and thyroid function soon adapt to the daily strain. Testosterone levels, however, were markedly decreased throughout the 20 days while LH levels remained unchanged. Thus i*** appears that the reproductive endocrine system is more susceptable to long‐term stress effects than the more vital functions of the thyroid and adrenal glands.
Obstruction of the tubuli recti and ductuli efferentes by dilated veins in the testes of men with varicocele and its possible role in causing atrophy of the seminiferous tubules
Authors: Nistal M. , Paniagua R. , Regadera J. , Santamaria L.
Published: May 2008

Hormone measurements, spermiograms and testicular biopsies studies were performed in young with varicocele. In addition, the testes and epididymides of 27 adults with varicocele were obtained from autopsies. Light and electron microscopic examination of biopsy and autopsy specimens revealed two types of lesions in testes with varicocele: 1) a diffuse lesion consisting of abnormal spermatozoa and spermatid morphology and sloughing of immature spermatozoa and spermatid: 2) focal lesion, distributed irregularly throughout the testicular parenchyma, affecting several small groups of seminiferous tubules. Each of these groups corresponded to a testicular lobule and showed different degrees of tubular atrophy, so that the focal lesions were distributed in a mosaic pattern. The testicular interstitium showed dilated veins and venules, and progressive collagenization. Some testes showed dilated veins in the rete testis, which compressed several tubuli recti and caused tubular atrophy in the seminifeous tubules opening into these tubuli recti. Other testes showed dilated young veins among the ductuli efferentes, and the rete testis channels appeared to be dilated. Among the different etiological mechanisms which have been suggested to for testicular lesions in varicocele. tubular obstruction at the level of either the tubuli recti or the ductuli efferentes might be responsible for lesions leading to testicular atrophy.
A modified method for collection of rete testis fluid from the rat
Authors: Yanwan Wu , Dong Yuan
Published: May 2008

Collection of rete testis fluid (RTF) from rats and monkeys has previously involved ligation of the efferent ducts (EDL) for 18–27 h before transcapsular cannulation of the swollen intratesticular rete (Tuck et al. 1970; Waites & Einer‐Jensen 1974). Our concern for possible contamination of such samples by lymph or capillary exudate prompted us to adopt a modified microcannulation procedure in order to reduce such contamination. Other methods for collecting RTF include cannulating ‘efferent ducts’ with or without previous EDL, respectively, in rabbits (Cooper et al. 1976) and rats (Free & Jaffe 1979). The latter cannulation technique proved difficult to repeat, and the volume of RTF collected was insufficient for analysis. This paper describes a method for routine collection of RTF from anaesthetised rats following cannulation of the extra‐testicular rete testis after efferent duct ligation.
Effects of short‐ and long‐term administration of tamoxifen on hCG‐induced testicular steroidogenesis in man: no evidence for an oestradiol‐induced steroidogenic lesion*
Authors: van Bergeijk L. , Gooren L. J. G. , van der Veen E. A. , de Vries C. P.
Published: May 2008

hCG‐induced testicular desensitization is characterized by inhibition at the level of the C‐17,20‐lyase enzyme. This defect has been attributed to an early rise in oestradiol (E2) following hCG administration. To test this hypothesis the E2‐receptor antagonist, tamoxifen, was employed. From in vitro studies the evidence suggests that tamoxifen depletes the E2‐receptor within 24 h. In this in vivo study, short‐term (36 h) administration of tamoxifen (to 6 eugonadal men) did not affect basal plasma levels of LH, FSH, 17α‐hydroxyprogesterone (17‐OHP), testosterone (T) and E2, whereas long‐term (3 months) tamoxifen with treatment of 6 normogonadotrophic oligozoospermic men increased LH and T levels, indicating a biological effect of tamoxifen. The response of 17‐OHP, T, E2 and the 17‐OHP/T ratio to hCG was similar in short‐term and long‐term tamoxifen‐treated men as well as in 6 untreated eugonadal male controls. These results do not suggest a role for endogenous E2 in the hCG‐induced testicular steroidogenic block.
Development of stage‐specific paracrine regulation of Leydig cells by the seminiferous tubules
Authors: Bergh Anders
Published: May 2008

The size of peritubular Leydig cells surrounding tubules in different stages of the spermatogenic cycle was determined in 43‐ and 47‐day‐old male rats. A stage‐dependent variation in the size of peritubular Leydig cells was not present in 43‐day‐old rats, but by 47 days those Leydig cells closely adjacent to tubules at stages VII–VIII were larger than others. At 43 days of age spermatogenesis had developed up to step 18 spermatids in late stage VI tubules. At 47 days of age the first mature sperm had just been released from the seminiferous epithelium, and consequently the first wave of the spermatogenic cycle was completed. Tubules at stages VII–VIII therefore acquire the ability to influence surrounding Leydig cells when they contain step 19 spermatids. It remains to be shown whether this maturation step is due to inherent maturation of the Sertoli cells or if step 19 spermatids specifically modulate Sertoli cell function.
Nuclear chromatin decondensation of spermatozoa in vitro: a method for evaluating the fertilizing ability of ovine semen
Authors: Rodriguez H. , Ohanian C. , Bustos‐Obregon E.
Published: May 2008

Spermatozoa obtained from testes, epididimydes and complete ejaculates of healthy rams during the breeding and non‐breeding seasons were induced to show nuclear chromatin decondensation by controlled exposure to dithiotreitol (DTT) and sodium dodecyl sulphate (SDS) in vitro. A gradual resistance to decondensation was shown by sperm during epididymal transit, confirming a progressive increase in the prevalence of chromatinic disulphide bonds during sperm maturation in this species. A high % of stable (non‐decondensed) sperm nuclei after treatment (79%) was found in semen from rams with normal fertility (64% non‐return rate at first oestrus). Opposite changes were found in the semen from rams having low fertility rates (37%), as these showed only 31% of stable sperm nuclei. There were no differences in the spermiograms of these two groups. When semen from the same rams was tested during the non‐breeding season, a similar relationship was found, although in both groups there was a higher % of sperm with stable nuclei than during the breeding season. The possible role of seminal plasma and of some of its constituents (e.g., zinc) on the decondensation of ram sperm nuclear chromatin was also studied. The presence of seminal plasma and the addition of zinc largely or completely inhibited the decondensation of ram sperm nuclear chromatin whilst the reverse situation was seen following the addition of ***
Dynamic study of serum gonadotrophin and testosterone levels in gossypol‐treated men Long term follow‐up study of 60 cases
Authors: Gui‐yuan Zhang , Bilian Xiao , Zhen‐wen Chen , Ji‐chuan Zhu , Guang‐dong Meng
Published: May 2008

The endocrine effects of gossypol were studied in 26 men for 52 months before, during and after treatment. A further 34 subjects were studied after cessation of gossypol treatment. A control group of 60 age‐matched volunteers were monitored for 1 year. No significant changes in testosterone levels were observed during the course of study. Serum LH levels were, however, significantly higher (P < 0.001) during earlier phases of gossypol treatment and returned to normal after cessation of treatment except in 14 men with persistent disruption of spermatogenesis. In the latter, serum levels of LH and testosterone were significantly higher (P < 0.001) than those found in normal subjects and significantly lower (P < 0.05) than in those subjects in which spermatogenesis had recovered. Serum FSH levels did not rise during the first 8–9 months of treatment with gossypol, although azoospermia generally occurred within 76 days of commencing treatment. After 9 months of treatment, serum FSH levels rose gradually and remained significantly elevated (P < 0.001) after cessation of treatment in both the azoospermic/oligospermic group and in the group in which spermatogenesis recovered. In the latter group the serum levels of FSH were correlated significantly with the sperm concentration.
Morphological factors influencing the penetration of human sperm into cervical mucus in vitro
Authors: Jeulin C. , Soumah A. , Jouannet P.
Published: May 2008

The efficiency of cervical mucus in filtering out single, multiple and associated abnormalities of human spermatozoa was determined. Twenty semen samples which gave a normal in vitro cervical mucus penetration test (CMPT) were analysed before and after migration using a detailed classification system (13 categories). The % of normal forms was significantly increased in cervical mucus (59.5 vs 33.2%), whereas the % of sperm with single, multiple or associated abnormalities of the midpiece or of the flagellum were found to decrease significantly in cervical mucus. Sperm with single or multiple abnormalities confined to the head migrated similarly to normal forms. The decrease in amorphous and elongated tapering sperm was explained by their more frequent association with other defects of the midpiece and/or of the flagellum.
Cross‐sectional study of semen parameters in a large group of normal Chinese men
Authors: Wang C. , Chan S. Y. W. , Leung A. , Ng R. P. , Ng M. , Tang L. C. H. , Ma H. K. , Tsoi W. L. , Kwan M.
Published: May 2008

Testicular size, semen characteristics and serum reproductive hormone levels were studied in 1239 normal Chinese men. The mean testicular size of 17 ml in this group of subjects was similar to other reports in Orientals but was smaller than that reported in Caucasians. Semen volume, and the sperm count, motility and morphology were comparable to those described for Caucasian subjects. The cervical mucus penetration test (Kremer's test) and the zona‐free hamster egg penetration test were also studied in some of the subjects. Age showed a positive correlation with the serum levels of FSH and LH, which were highly correlated with each other. Sperm concentration and the total sperm count were positively correlated with testicular volume, the length of abstinence, the % of sperm with an oval head, and the zona‐free hamster egg penetration test, but were negatively correlated with serum FSH. Sperm motility was negatively correlated with the duration of abstinence but was positively correlated with the % normal sperm. The results constitute a normal data base from Chinese men for comparison with studies in male infertility and fertility regulation.
Acrosome loss in human sperm incubated in vitro under capacitating conditions
Authors: Mallett Philippa J. , Stock Caroline E. , Fraser Lynn R.
Published: May 2008

Human sperm were incubated under capacitating conditions and, at selected points up to 24 h of incubation, motile cells were fixed and assessed with the electron microscope for the presence or absence of the acrosome. Two methods of sample preparation were compared. In the first, semen samples were washed, incubated and filtered through glass beads to select motile cells before fixation. In the second, motile sperm were allowed to swim up into medium; this produced >95% motile cells which were then incubated and fixed as required. In both series of experiments a significant increase in acrosome loss with time was observed (P= 0.00013), although only 10.5% of cells had lost the acrosome after 24 h. It is concluded that overt acrosome loss occurs less frequently in human sperm than in those of commonly used laboratory animals.
The importance of the Leydig cells in the vascular response to hCG in the rat testis
Authors: Setchell B. P. , Rommerts F. F. G.
Published: May 2008

The increase in permeability of the testicular blood vessels following an injection of hCG into rats is abolished completely if the animals are treated 3 days earlier with ethane dimethane sulphonate (EDS), a compound that effectively eliminates Leydig cells from the testes. As there is other evidence that androgens or prostaglandins are not involved in this vascular response, further studies will be necessary to determine whether these data mean that another vasoactive substance is secreted by the Leydig cells or whether the EDS also eliminates other cells besides the Leydig cells, for example the mast cells found in the vicinity of the testicular artery.
Increase in progressive motility and improved morphology of human spermatozoa following their migration through Percoll gradients
Authors: Pousette Åke , Åkerlöf Eva , Rosenborg Lennart , Fredricsson Bengt
Published: May 2008

Human spermatozoa were separated on the basis of their motility in a discontinous Percoll‐gradient made up in tissue culture medium containing 10% (v/v) human serum (TCMS). Portions of ejaculates were placed on top of the gradients. After 3 h at 37°C the bottom 1.5 ml was collected and the sperm washed free of the Percoll solution by centrifigation at 240 × g after dilution in TCMS. In this way the spermatozoa were separated from seminal fluid by means of the swimming rate of the sperm. When semen samples from normal men were used, total recovery of sperm after separation on a Percoll gradient was 21 ± 2.3%. The progressive motility index increased by a factor of 15 ± 1 when comparing separated samples with the same unseparated ejaculate, and the frequency of sperm with normal morphology increased from 60 to 85%. The improvements in these semen samples was attributable to the Percoll separation as the washing procedure itself was without effect. Using this method sperm of relatively unifirm motility and morphology can be collected. These may then be used for further biochemical and physiological studies. Improved sperm quality was also obtained when samples from patients with abnormal semen profiles were separated in this way, although the degree of improvement was much more variable than that obtained with semen from normal fertile men. This indicates that this method can be used in clinical practice in selected cases for the preparation of sperm for insemination or for in‐vitro fertilization.
The effect of doxycycline in infertile couples with male accessory gland infection: a double blind prospective study
Authors: Comhaire F. H. , Rowe P. J. , Farley T. M. M.
Published: May 2008

Male accessory gland infection (MAGI, epididymo‐prostato‐vesiculitis) with abnormal semen quality was rarely the only abnormality in infertile couples since it occurred in no more than 1.6% of 2871 couples evaluated in 7 centres during a 3‐year period. Both partners of 33 infertile couples with no other demonstrable abnormality than abnormal semen and MAGI consented to participate in a double blind trial and were treated with either doxycycline. 100 mg/day for 1 month (20 couples) or placebo (13 couples). Follow‐up during a total of 175 couple‐months included semen analysis and the recording of pregnancy. Pregnancy occurred in 2 of the doxycycline‐treated couples (10%) and in 1 of the placebo treated couples (8%), corresponding with conception lates per month of 1.9% and 1.5%, respectively. Sperm motility and, to a lesser extent, morphology showed improvement in both groups. Evidence of infection, namely increased numbers of white blood cells and positive sperm culture, disappeared in both the doxycycline‐treated and placebo group. It is concluded that features of MAGI in semen may regress spontaneously and are not influenced by the doxycycline treatment. The concomitant improvement of sperm motility and morphology still does not seem to enhance the probability of conception.
Immunohistochemical and ultrastructural study of Sertoli cells in androgen insensitivity
Authors: Aumüller G. , Peter St.
Published: May 2008

The ultrastructure of a perfusion‐fixed gonad in a case of androgen insensitivity was studied using thin sections and freeze‐fracture replicas. The distribution and arrangement of intermediate filaments in Sertoli cells was visualized immunohistochemically using an antibody against vimentin. Leydig cells lacked Reinke crystals, but contained all of the cytoplasmic organelles involved in steroid synthesis and additionally several lysosomes. The basement membrane and the basal lamina of the testicular tubules were considerably thickened. The testicular tubules consisted of gonocytes and Sertoli cells which had an immature nuclear structure, incomplete development of intercellular junctions and a primitive distribution pattern of intermediate cytoplasmic filaments. The previously reported differences in electron density of Sertoli cell cytoplasm are a non‐specific feature without significance to Sertoh cell maturation.
Biochemical and histochemical studies on the pituitary‐testicular axis in obese (C57B1/6J‐ob/ob) mice
Authors: Kühn‐Velten N. , Codjambopoulo P. , Haider S. G. , Passia D. , Kley H. K. , Herberg L. , Staib W. , Goslar H. G.
Published: May 2008

Male C57B1/6J‐ob/ob mice (4 months old) and their homozygous lean controls were compared with respect to pituitary LH secretion and functional parameters of purified Leydig cells in vitro. Compared with controls, obese mice showed reductions in the following parameters: Plasma testosterone levels (reduced by 57%), hCG‐stimulated testosterone formation in vitro (by 31%), conversion of progesterone to androgens by Leydig cells (by 39%), and GnRH‐stimulated LH secretion (by 26%). Lipid accumulation and a 37% decrease in naphthylesterase activity in the Leydig cells as well as hyperplasia of pituitary gonadotrophs were observed histochemically in obese mice. The changes in testicular endocrine function in obese mice are interpreted as consequences of pituitary dysfunction.
Spermatogenesis in Snell dwarf, little and congenitally hypothyroid mice
Authors: Matsushima Masahiro , Kuroda Kanami , Shirai Masafumi , Ando Ko , Sugisaki Tetsuro , Noguchi Tetsuya
Published: May 2008

The status of spermatogenesis in Snell dwarf, little and congenitally hypothyroid mice was studied. In all of these mice with a hormone deficiency the seminiferous tubules were smaller in size and contained fewer spermatogonia, spermatocytes, spermatids and spermatozoa than did those of normal control mice. There was no substantial difference in the Johnsen score between the hormone‐deficient mice and normal control mice, but the former had underdeveloped seminiferous tubules with a corresponding paucity of germ cells, which may be partly responsible for the infertility of these mice. In the present study, growth hormone and thyroxine were administered separately to growth hormone‐deficient and thyroxine‐deficient mice, respectively. Such replacement therapy brought about an increase in cell counts of the seminiferous tubules and in sperm counts in both groups.
The fate of acrosomal staining during the acrosome reaction of human spermatozoa as revealed by a monoclonal antibody and PNA‐lectin
Authors: Kallajoki M. , Virtanen I. , Suominen J.
Published: May 2008

A monoclonal anti‐human sperm antibody, raised against an acrosomal antigen and indicated to recognize in boar sperm the serine protease, acrosin, stained in human spermatozoa a 50 Kd antigen and several others in the region 24–34 Kd by immunoblotting. The 50 Kd band and the region of 30–34 Kd showed proteolytic activity by zymographic enzyme detection. The fate of the antigen was studied in the acrosome reaction induced by the calcium ionophore A23187. In control incubations 69.5 ± 14.2% (mean ± SD) of the spermatozoa had intact acrosomal staining according to indirect immunofluorescence using this antibody whereas in acrosome‐reacted samples only 21.0 ± 2.0% of the sperm were stained. Another marker for the acrosome, peanut agglutinin‐lectin (PNA), was used to detect the acrosome with similar results. Acrosome reactions were verified by electron microscopy. The present results indicate that the corresponding antigen, evidently acrosin, and PNA‐positive material are liberated during the acrosome reaction which suggests that they are not bound to the inner acrosomal membrane but are components of the acrosomal matrix.
On the use of adenosine triphosphate for estimation of motility and energy status in human spermatozoa
Authors: Pousette Åke , Åkerlöf Eva , Lundin Arne , Rosenborg Lennart , Fredricsson Bengt
Published: May 2008

Following the development of a reliable technique for analysis of total ATP in semen, the ATP concentration of untreated human ejaculates was compared with progressive sperm motility during 3 continuous days following ejaculation. As correlation was poor between these variables, sprermatozoa were separated by self‐migration on Percoll gradients and washed free of Percoll solution. ATP concentration and sperm motility was then assessed. Aliquots of separated sperms were transferred to separate tubes, and increasing concentrations of sera were added prior to incubation at 37°C for 4 days. Under these conditions a good correlation between specific progressive motility and specific ATP content was found (r = 0.88). A high concentration of human blood serum (10% v/v) was found to be necessary for maintainance of motility, but neither the addition of bovine serum albumin nor an increase in the glucose concentration had any effect on sperm motility or on the ATP concentration. Addition of different fractions of human blood serum indicated that the components mainly responsible for maintaining sperm motility are > 10 000 daltons. Our findings strongly indicate that ATP assessment can be used to estimate the energetic condition of a human sperm population after its separation from seminal plasma. It is suggested that this in vitro system can be used to define optimal conditions for sperm function.
Large forms of immunoreactive calcitonin in human seminal fluid exhibit PDN‐21 immunoreactivity
Authors: Bucht Elisabet , Arver Stefan , Sjöberg Hans Erik
Published: May 2008

Semen samples were obtained from 4 healthy males on two occasions. One of the samples was liquefied at room temperature for 20 min while the other was diluted immediately with buffer to suppress proteolysis. A pool of immunoextracted calcitonin from liquefied and diluted samples, respectively, was subjected to gel chromatography in a separate experiment. In lyophilized samples the majority of immunoreactive calcitonin (CT) had an approximate molecular weight (Mr) of 10 kilodaltons (Kd) and immunoreactive PDN‐21 was also present in the fractions. In diluted ejacultes CT was composed of larger species as well as the 10 Kd form. PDN‐21 was present in fractions containing the 10 Kd form but could not be detected in the peaks of larger molecular weight, probably because of lower sensitivity of the PDN‐21 assay compared to the CT assay. Our finding of CT and PDN‐21 in the same fractions after immunoextraction with CT antibodies suggests the presence of proforms of CT in human seminal fluid.
Temporary inhibition of the initiation of motility of demembranated hamster sperm by high concentrations of ATP
Authors: Yeung C. H.
Published: May 2008

ATP is obligatory for sperm movement. When sperm from the hamster cauda epididymis were washed and exposed to a demembranation and re‐activation medium containing 0.5 or 1 mM ATP, the maximal % motility was attained almost immediately. Increasing the ATP concentration to 3 mM caused a 5 min delay in development of maximal re‐activation and a change in the beating pattern as indicated by a decrease in the % of forward progressing sperm, decreased linear velocities but unchanged curvilinear velocities. The inhibition of motility initiation by 3 mM ATP was abolished either by the inclusion of trypsin (50 ng/ml) or by a brief pre‐incubation of the sperm in the demembranation and reactivation medium before the addition of ATP. However, the beneficial effect of pre‐incubation was prevented if trypsin inhibitor was present. This indicates that the inhibition by ATP acts via a trypsin‐sensitive regulatory system, and suggests that an endogenous protease activity is necessary to overcome this. Re‐activation of sperm from the caput epididymis by all three concentrations of ATP showed similar lag periods which could also be diminished by pre‐incubation or trypsin. The results suggest that, in addition to developing their motile capacity during maturation in the epididymis, sperm exonemes change their sensitivity to ATP concentrations.
Inhibition of RNA and DNA synthesis in Sertoli cells by co‐culture with spermatogenic cells
Authors: Rivarola M. A. , Sanchez P. , Saez J. M.
Published: May 2008

Sertoli cell monolayers were prepared from 30‐day‐old rat testes and cultured for 7 days to eliminate contaminant germ cells. Some of these monolayers were co‐cultured with a spermatogenic cell preparation enriched in pachytene spermatocytes and round spermatids (fraction 3 from a Percoll gradient), isolated from 30‐day‐old rat testes. After co‐culture for 4 to 48 h, germ cells were removed. RNA synthetic activity in rat Sertoli cell cultures alone was 216 800 ± 66 480 dpm [3]uridine.2h−1.106 cells−1 (mean ± SD) compared to 98 390 ± 23 595 in rat Sertoli cells which had been co‐cultured with germ cells of fraction 3 for 24 h (P < 0.01). By contrast, RNA synthesis in Sertoli cell monolayers prepared from immature pigs were unaffected by co‐culture with rat germ cells. A similar inhibitory effect of germ cells was observed in rat Sertoli cells stimulated with FSH or testosterone. Culture medium, conditioned by 20 h culture of a fresh preparation of rat spermatogenic cells of fraction 3, was active in inducing the inhibitory effect on RNA synthesis in rat Sertoli cells. Co‐culture of rat Sertoli cells with germ cells of this fraction also decreased the incorporative of [3H]thymidine into DNA in rat Sertoli cells, from 9061 ± 3339 to 4766 ± 526 dpm.2h−1.106 cells−1 (P < 0.01), but no such change was found in pig Sertoli cells. A different spermatogenic cell preparation, partially deprived of pachytene spermatocytes (fraction 5), stimulated rat Sertoli cell DNA synthesis (Sertoli alone 7833 ± 2550, Sertoli cells which had been in co‐culture with germ cells of fraction 5, 13 300 ± 2279 dpm.2h−1.106 cells−1, P < 0.05). These inhibitory actions of some germ cells on Sertoli cells were observed together with the previously reported simultaneous stimulatory effect of Sertoli cells on germ cells. These Sertoli cell‐germ cell interactions of detected in culture may represent regulatory influences operating in vivo.
Effect of protein malnutrition on glycoprotein synthesis by testes of 20‐day‐old rats
Authors: Guaragna Regina , Rosa Neila B. S. , Moraes Galba G. S. , Perry Marcos L. S. , Bernard Elena A.
Published: May 2008

The effects of protein malnutrition during the nursing period on glycoprotein biosynthesis by testes of 20‐day‐old rats was studied. Pregnant Wistar rats were housed individually. On the day of delivery they were divided into two groups: one was fed a control diet (25% casein) and the other a low‐protein diet (8% casein) for a period of 20 days. Body and testis weights of pups suckled by the malnourished mothers were significantly lower than those of the pups suckled by normally‐fed mothers. The seminiferous tubules o malnourished rats showed a significant decrease in diameter and in the stage of development of spermatogenesis. Whole testes of normally‐fed 20‐day‐old rats showed significantly greater [2‐3H]mannose incorporation into glycoproteins than did the testes of malnourished rats of the same age. The microsomes of normally‐fed rats showed significantly higher GDP: mannose polyprenyl mannosyl transferase activity than did microsomes from malnourished rats, and this difference increased when exogenous dolichyl‐phosphate was added to the incubation medium. These results indicate that protein malnutrition decreases GDP: mannose polyprenyl mannosyl transferase activity in the microsomes of testes from 20‐day‐old rats.
Immunohistochemical demonstration of placental alkaline phosphatase in various states of testicular development and in germ cell tumours
Authors: HUSTIN J. , COLLETTE J. , FRANCHIMONT P.
Published: May 2008

Immunohistochemical localization of placental alkaline phosphatase (PlAP) has been performed on eighty‐two samples of normal (embryonic, fetal, infantile or adult), cryptorchid or tumorous testicular tissue. The isoenzyme could be demonstrated at the cell membrane of primitive, embryonic germ cells but not in other normal tissues. In‐situ carcinomas and seminomas were positively stained in 93% and 94% of cases respectively. The percentage of positivity decreased with tumour differentiation. It is suggested that PlAP is a marker of primitive germ cells and that it reappears in germ cell neoplasms by gene derepression.
The function of the primordial germ cell in extragonadal tissues
Authors: FRIEDMAN N. B.
Published: May 2008

The identity of the germ cell tumours of the pineal and the thymus with those of the testis and ovary suggests that the widely disseminated primordial germ cells might subserve some special function in these sanctuaries. It is proposed that thymic localization might be required for the conveyance of genetic haematological and immunological information and that the pineal–diencephalic localization could programme neuroophthalmic tissues prior to the development of the blood–brain barrier. The latter speculation was tested by producing allergic encephalomyelitis in thymectomized, bursectomized and thymobursectomized chickens. It was found that although thymectomy and, to a lesser extent, bursectomy decreased the severity of the experimental encephalomyelitis the combined procedure resulted in more severe inflammatory lesions. This may be due to release of suppressed intraneural immunological mechanisms in the somatically impaired bird.
Cell lines of human germinal cancer
Authors: CASPER J. , SCHMOLL H. ‐J. , SCHNAIDT U. , FONATSCH C.
Published: May 2008

Three new established human germ cell tumour lines, H 12.1, H 12.5 and H 12.7, are described. Cytogenetic and growth characteristics, morphology, histology, and tumour marker and steroid hormone production in vitro and/or in vivo revealed properties commonly found in other germ cell tumour lines and in patients with these tumours. In vitro oestrone and oestradiol were produced by the H 12 lines and four other lines (833 KE, 1156 Q, 1428 A and 2102 EP) under high and low density (differentiation inducing) conditions. AFP was produced by one line and β‐hCG by six of seven lines under low density conditions. The pattern of oestrogen production suggests that these hormones could be useful in AFP and β‐hCG negative patients. Differentiated elements of somatic and extraembryonic character, observed in tumours of H 12.1 and H 12.7, underline the value of these lines in the study of differentiation and other germ cell tumour related questions.
Analysis of the autoimmune response in an ‘in situ’ carcinoma of the testis
Authors: LEHMANN D. , MÜLLER Hj.
Published: May 2008

Immune response in a patient with an in‐situ seminoma of the testis was studied. Immunohistochemical examination of the testicular biopsy demonstrated remarkable intracellular and membraneous accumulation of IgG antibodies in the atypical spermatogonia. Using FITC‐conjugated Concanavalin A, an abnormally high binding of Concanavalin A was detected in the transformed cells. Circulating antibodies directed against an antigenic determinant (60 kD) which is expressed on normal human spermatozoa and lymphocytes were found only in the patient's serum and not in 500 control sera. The role and possible diagnostic value of this autoimmune reactivity in testicular malignancies are discussed.
Management of carcinoma‐in‐situ of the testis
Authors: MAASE H. VON DER , GIWERCMAN A. , MÜLLER J. , SKAKKEBÆK N. E.
Published: May 2008

Carcinoma‐in‐situ (CIS) of the testis progresses to invasive cancer within 5 years in 50% of cases, and therefore requires therapeutic intervention. CIS is probably eradicated by intensive cancer chemotherapy but this is too toxic for the management of non‐invasive disease. Eight patients with unilateral testicular cancer and contralateral CIS received localized irradiation (20 Gy in ten fractions of 2 Gy) to the remaining testis: after 3 months the CIS cells had disappeared and ‘Sertoli‐cell‐only’ tubules were found. LH and FSH levels were elevated but testosterone levels remained fairly constant. Localized irradiation should be considered as the treatment of CIS in the contralateral testis of testicular tumour patients unless chemotherapy is indicated for the primary tumour.

Unilateral orchidectomy is recommended for unilateral CIS associated with infertility or testicular maldescent. Localized testicular irradiation should now be considered for bilateral disease. Patients with the androgen insensitivity syndrome should normally be treated with bilateral orchidectomy, but irradiation may be useful in selected cases.
Prognostic factors in metastatic testicular cancer
Authors: VOGELZANG N. J.
Published: May 2008

The prognosis for metastatic testicular cancer has been analysed in seven reports using multivariate analysis. Serum hCG is the most important factor. Volume of metastases, serum LDH and serum AFP are also of prognostic value. Bone, liver, nodal or retroperitoneal metastases are not independent prognostic factors. The prognosis for extragonadal nonseminomas remains in dispute. Future studies should categorize poor prognosis patients using one of the several available prognostic formulas. No consensus yet exists on optimal treatment for such patients.