bluuebunny/crossref_metadata_2025 huggingface dataset:
Localization of lactoferrin in the male reproductive tract
Authors: WICHMANN L. , VAALASTI A. , VAALASTI T. , TUOHIMAA P.
Published: May 2008

The immunohistochemical localization of lactoferrin in the normal human prostate, seminal vesicle, vas deferens, epididymis and testis was studied using the per‐oxidase‐antiperoxidase method at the light and electron microscopical level. Lactoferrin immunoreactivity was localized in the glandular epithelial cells and granulocytes in the prostate and seminal vesicle. In the prostate, lactoferrin showed an uneven distribution; some of the glands contained exclusively positive cells and others were completely lactoferrin netative, while the rest contained scattered positive cells. The seminal vesicles were divided into three segments, and their lactoferrin content varied significantly although it was always epithelial. The ductus deferens, epididymis and testis contained no lactoferrin. In conclusion, lactoferrin was found in the prostate and seminal vesicles, but not in the testis.
A simple spot test for the detection of fructose deficiency in semen
Authors: JUNGREIS E. , NECHAMA M. , PAZ G. , HOMONNAI T.
Published: May 2008

The concentration of fructose in seminal plasma is an important variable that should be measured in the assessment of fertility status. The current method requires the boiling of diluted seminal plasma with resorcinol. The present study offers a new two‐point spot test which uses the metol method. The results of analysis of 104 seminal plasma samples are in excellent correlation with the resorcinol method. The reported method is easy to use, inexpensive, and can be introduced into any laboratory.
Pulsatile LHRH therapy in patients with oligozoospermia and disturbed LH pulsatility
Authors: AULITZKY W. , FRICK J. , HADZISELIMOVIC F.
Published: May 2008

Pulsatile administration of LHRH can drive the pituitary to secrete LH and FSH in a pattern that closely mimics the physiological pattern of the hypothalamic‐pituitary‐gonadal axis. As there is evidence that infertility in some men is due to dysfunction of this axis, 14 men with reported infertility of more than 2 years duration were treated by long‐term pulsatile LHRH therapy. They were 24–42 years of age, with variable degrees of oligozoospermia, elevated FSH levels but normal LH and testosterone levels. The number of endogenous LH pulsed/24 h was less than eight in all 14 subjects. The degree of testicular damage was assessed on semi‐thin sections prepared from biopsies of both testes. Scores for spermatogonia per seminiferous tubule (SPT) were calculated from the actual number of Ad‐spermatogonia/tubule. Patients were grouped according to sperm density and SPT score (groups I—III). Pulsatile LHRH therapy was administered by means of a portable infusion pump; 4 μg LHRH were administered subcutaneously every 120 min for a period of 6 months. This treatment restored the normal pattern of LH secretion to 12 LH pulsed/24 h in all patients. A statistically significant decrease of mean FSH levels to normal, and an increase of mean LH levels was observed in most of the 14 patients. Testosterone values did not change in any group. Marked improvement of the sperm count was observed in eight out of 14 patients (groups I and II) and three pregnancies were reported during the treatment periods. These results suggest that some states of male infertility are due to hormonal dysregulation and that these patients may benefit from pulsatile LHRH therapy.
Inhibin secretion is influenced by Ley dig cells: evidence from studies using the cytotoxin ethane dimethane sulphonate (EDS)
Authors: KRETSER D. M. DE , O'LEARY P. C. , IRBY D. C. , RISBRIDGER G. P.
Published: May 2008

Adult male rats given a single intraperitoneal injection of the Leydig cell cytotoxin ethane dimethane sulphonate (EDS) show a significant decrease in testosterone from 7 to 14 days, and elevation of serum FSH and LH levels commencing 7 days after treatment, returning to normal at 28 days for LH and 49 days for FSH. A significant rise in serum inhibin levels was seen at day 14 after EDS treatment with levels returning to normal at day 49. In a second series of experiments, silastic implants of testosterone, either 2.5 cm or 22.5 cm in length, were introduced subcutaneously into adult male rats which were treated with EDS 10 days later. Both doses of testosterone suppressed basal LH levels but did not significantly change FSH levels. The rise in FSH and LH levels seen in normal rats after EDS treatment did not occur in either group of testosterone‐implanted rats. However, serum inhibin levels rose significantly in both groups after EDS treatment, suggesting that the rise in serum inhibin levels was not due to stimulation arising from the increase in FSH levels after EDS treatment. The data suggest that the rise in serum inhibin levels after EDS treatment is linked to destruction of the Leydig cells through mechanisms that require further investigation.
Evaluation of the relative importance of endocrine and paracrine factors in control of the levels of inhibin in testicular interstitial fluid
Authors: SHARPE R. M. , MADDOCKS S.
Published: May 2008

This study aimed to identify the role of endocrine (FSH, LH, testosterone) or paracrine (Leydig or germ cell) factors in control of the secretion of inhibin into testicular interstitial fluid (IF). This was done by measuring inhibin and testosterone levels in IF, and serum gonadotrophin and testosterone levels in adult rats following the destruction of Leydig cells with ethane dimethane sulphonate (EDS), alone or in combination with testosterone ester (TE) supplementation at various doses initiated at various times after EDS treatment. The effect of germ cell loss (induced by local testicular heating) on its own or in combination with the above treatments was also assessed.

Treatment with EDS led to major increases in the levels of inhibin in IF and of FSH and LH in serum whilst testosterone levels in IF and serum fell to undetectable levels. Supplementation with TE (1–25 mg) for 21 days from the time of EDS treatment failed to prevent the initial (+3 days) increase in IF levels of inhibin but thereafter suppressed inhibin to control levels or lower and grossly suppressed FSH and LH levels, irrespective of whether the dose of TE administered did (25 or 5 mg) or did not (1 mg) prevent major seminiferous tubule damage. Partial regeneration of Leydig cells and normalization of testosterone levels occurred in rats 21 days after treatment with EDS alone but this failed to normalize inhibin and gonadotrophin levels. When supplementation with TE (25 mg) was initiated at 3, 6 or 9 days after EDS treatment, IF levels of inhibin were normalized within 3 days and maintained thereafter in parallel with suppression of serum FSH and LH to below control levels. Seminiferous tubule damage induced by local testicular heating (43°C for 30 min) led to increased IF levels of inhibin 3 and 14 days later, in parallel with increased serum levels of FSH (but not LH). Suppression of FSH to subnormal levels in heat‐exposed rats by TE treatment (25 mg) restored IF inhibin to control levels or below, a change which still occurred when Leydig cells were destroyed by EDS treatment.

It is concluded that secretion of inhibin via the base of the Sertoli cell into testicular IF is controlled primarily by FSH, although local factors may play a minor role. These findings have important implications regarding the possible paracrine role(s) of inhibin in IF during puberty and in the normal adult testis.
Interaction of hCG with testicular interstitial fluid produces a leucotactic factor
Authors: VEIJOLA M. , RAJANIEMI H.
Published: May 2008

An intratesticular injection of hCG (5 ng) mixed with testicular interstitial fluid (IF) increases vascular permeability in the rat testis. The present results show that the permeability increase induced by this treatment is accompanied by a massive accumulation of polymorphonuclear leucocytes (PMNs) in both the testicular post‐capillary venules and the interstitium. Depletion of neutrophils in the circulation by treatment with anti‐neutrophil serum significantly inhibited the permeability increase induced by this treatment. An intratesticular injection of PMNs (107 cells) or hCG alone had no effect on permeability, but a combination of the two caused a significant increase in permeability. The PMNs were found to secrete a component in vitro which, when injected intratesticularly together with hCG, caused a permeability increase and a simultaneous massive accumulation of PMNs in the postcapillary venules and interstitium. This permeability increase was prevented by the serine protease inhibitor p‐aminobenzamidine, suggesting an involvement of the plasminogen activator system in the response. The results suggest that hCG interacts with an IF component to produce leucotactic factors that increase permeability indirectly by attracting PMNs to the tissue, and that the IF component may originate in the PMNs.
Computer‐assisted image analysis of sperm concentration in human semen before and after swim‐up separation: comparison with assessment by haemocytometer
Authors: CHAN S. Y. W. , WANG C. , SONG B. L. , LO T. , LEUNG A. , TSOI W. L. , LEUNG J.
Published: May 2008

Evaluation of male fertility is based predominantly on results from semen analysis and determination of the sperm concentration is one of the main parameters of the analysis. The availability of a fully automated videomicrographic digital image analyser would offer both an objective and rapid method for determination of the sperm concentration. In the present study the sperm concentration in 327 semen samples was determined by haemocytometer according to the World Health Organization guidelines, and also by a computer‐assisted digital image analyser system. Results were classified according to the routine procedure (haemocytometer) before statistical analyses. The computerized measurements caused a shift to the right in the frequency distribution of sperm concentration. Sperm concentrations were more often overestimated significantly (P < 0.001) by the computerized measurements in semen samples with concentrations up to 80.0 × 106/ml. This overestimation seemed to be caused by the presence of particles in seminal plasma that were recognized incorrectly as sperm by the computer program. The computerized digital image analyser gave an average sperm concentration of 2.2 ± 0.6 × 106/ml (mean ± SEM) in 17 azoospermic semen samples while the routine procedure did not detect the presence of sperm cells. After removing the seminal plasma by washing and centrifugation with culture medium, and using the swim‐up procedure to harvest motile sperm, the computerized measurements showed comparable results with the routine procedure for those sperm preparations (n= 44) with sperm concentrations >5.0 × 106/ml. In sperm preparations with concentrations < 5.0 × 106/ml (n= 75) there was still a significant (P < 0.001) overestimation of the sperm concentration. These findings indicate that the CellSoft computer program has to be improved to allow correction for the particles present in seminal plasma which are not sperm. For determination of the sperm concentration in seminal plasma‐free preparations, an effective method to remove these particles is also necessary to obtain reliable results with the present computer‐assisted digital image analyser system.
A novel maturation marker for human Sertoli cells
Authors: BAUMAL R. , BAILEY D. , GIWERCMAN A. , SKAKKEBAEK N. , STRATIS M. , MARKS A.
Published: May 2008

The maturation of Sertoli cells at puberty is critical for the initiation and maintenance of spermatogenesis. Little is known about the factors which control Sertoli cell maturation. We have examined Sertoli cells in testicular specimens from 44 subjects, ranging in age from 6 months to 67 years, for reactivity with a mouse monoclonal antibody (M2A) using the immunoperoxidase reaction. We found positive reactivity with prepubertal but not postpubertal Sertoli cells, suggesting that the antigen defined by M2A was a marker of immature Sertoli cells. This marker may be useful for studying the factors which influence Sertoli cell maturation during development of the testis.
Bias to routine semen analysis by uncontrolled changes in laboratory environment—detection by long‐term sampling of monthly means for quality control
Authors: KNUTH U. A. , NEUWINGER J. , NIESCHLAG E.
Published: May 2008

To detect systematic bias in the results of routine semen analysis over time, monthly means of semen parameters determined by the recommended WHO methods were computed. The analysis was based on a total sample size of 1784 ejaculates and included 18 months of observation. In addition to slight changes of morphology estimates caused by a change of laboratory staff, a major bias in the measurement of sperm motility could be detected. This observation triggered a search for changes in protocols not previously given the required attention. It revealed that the newly introduced use of polypropylene syringes with a mounted needle for accurate measurement of seminal volume impaired sperm motility. More detailed investigation by computerized sperm motion analysis in 10 semen samples treated simultaneously in different ways revealed that predominantly it was the needle which caused the drop in proportion of motile sperm (glass cylinder: 50.3 ± 4.1% vs. syringe + needle: 26.6 ± 5.3%; mean f SEM) and not the contact with the plastic material alone (syringe alone: 43.4 ± 4.8%). Other motion parameters such as curvilinear velocity (36.0 ± 1.6 μm/sec), linearity (78.5 ± 8.4%) and lateral head displacement (3.8 ± 0.9) were not influenced by the different methods of handling. The results indicate that long‐term sampling of monthly means may serve as part of a quality control scheme in semen analysis.
Objective measurement of testicular volume by ultrasonography: evaluation of the technique and comparison with orchidometer estimates
Authors: BEHRE H. M. , NASHAN DOROTHÉE , NIESCHLAG E.
Published: May 2008

Ultrasonography of the scrotum is a valid and safe method for differentiating testicular and extratesticular disease. In the present study the accuracy and reproducibility of ultrasonographic estimation of testicular volume was evaluated. A high correlation (r= 0.992) between actual testicular volume (measured by weighing and water displacement) and the volume determined by ultrasonography was found on examination of 14 testes from patients at autopsy without any systematic under‐ or over‐estimation. The ultrasound method proved to be highly reproducible with a coefficient of variation of 7.0 ± 0.7%. Using different ultrasound scanners did not significantly influence the results obtained. The comparison of Prader orchidometer measurements, performed by four different clinical investigators, and ultrasonography in 256 patients revealed a significant correlation of 0.91, but the degree of correlation was dependent on the investigator's clinical experience. Volume estimates by comparative palpation appear to be valuable in most clinical settings. Ultrasonography provides an excellent tool for determining testicular volume when objective, accurate and reproducible measurements of testicular volume are required.
Use of laparoscopy in children with impalpable testes
Authors: DOIG CAROLINE M.
Published: May 2008

Our experience in Manchester using laparoscopy in children with impalpable testes is detailed. Eighty‐seven testes were investigated in 77 patients. Of these, 36% had no testes, although in 28% evidence of a vas was found. In those testes which were found, surgery was performed, either orchidopexy or, if positioned high, microsurgery. Orchidectomy was rarely carried out (5 testes only).
Age‐dependent Sertoli cell responsiveness to germ cells in vitro
Authors: CASTELLÓN E. , JANECKI A. , STEINBERGER ANNA
Published: May 2008

This study investigated the effect of germ cells (>80% mid‐ and late‐pachytene spermatocytes) on the secretion of androgen binding protein (ABP) and transferrin by monolayer cultures of Sertoli cells isolated from rats aged 10, 18 or 26 days. There was an age‐dependent increase in secretion of ABP and transferrin. Treatment of the Sertoli cell monolayers with hypotonic buffer to remove residual germ cells reduced this increase significantly. On the other hand, addition of germ cells to hypotonic‐treated Sertoli cell monolayers increased both basal and FSH + testosterone‐stimulated ABP and transferrin secretion at all three ages, although Sertoli cells from 10‐day‐old animals showed the greatest response. Moreover, addition of germ cells reduced responsiveness to FSH + testosterone in Sertoli cell monolayers obtained from rats aged 18 or 26 days. In monolayers obtained from 10‐day‐old rats, the opposite effect was noted in the case of ABP secretion. The stimulatory effect of germ cells on ABP and transferrin secretion was proportional to their number, and was reversed 48 h after the germ cells added previously were removed by hypotonic treatment. Whereas the reversal was complete with cultures of Sertoli cells isolated from 18‐ and 26‐day‐old rats, approximately 40% of the stimulatory effect remained after removal of germ cells from cultures from the 10‐day‐old age group. Adhesion of germ cells to Sertoli cell monolayers was also found to be age‐dependent, with the largest proportion of added germ cells adhering to Sertoli cells isolated at 18 and 26 days of age. It is concluded that germ cells can significantly and differentially modulate the basal and hormone‐stimulated secretory activity of Sertoli cells in vitro and that Sertoli cell responsiveness to germ cells (pachytene spermatocytes) is age‐dependent and seems to appear early during the maturation process, before these germ cells appear in the testis.
Induction of infertility in female rats after active immunization with 24 kD antigens from rat testes
Authors: SHAHA C. , SURI A. , TALWAR G. P.
Published: May 2008

A unique polyvalent antiserum against whole washed human sperm was used previously to identify groups of antigens on spermatozoa. The antiserum, designated as Antiserum I, recognized the acrosome on sperm from several species and on Western blots recognized a 40 kD antigen in human sperm extracts. Antiserum I caused agglutination of human sperm and prevented interaction of mouse sperm and oocytes. This serum also recognized a band of 24 kD in rat testicular cytosol. In the present study this group of 24 kD proteins was used as an antigen preparation to actively immunize female rats. Immunization was carried out with two different adjuvants: nor‐muramyl dipeptide and SPLPS (a thyalated derivative of lipopoly‐saccharide). Both groups of animals showed significant antibody titres as detected by indirect immunofluorescence and by sperm agglutination tests. In both groups over 80% of the animals remained infertile, compared to 13% of the controls. It is concluded that a group of antigens in rat testes recognized by Antiserum I offer promise as candidates for a contraceptive vaccine.
Evaluation of the Hamilton‐Thorn 2030 motility analyser for routine semen analysis in an infertility clinic
Authors: NEUWINGER J. , KNUTH U. A. , NIESCHLAG E.
Published: May 2008

In routine semen analysis of 242 patients the values of sperm concentration, sperm motility and progressive sperm motility were measured with the Hamilton‐Thorn semen analyser (HT) and compared to the data obtained by conventional semen analysis according to the guidelines of the WHO. Overall, the HT gave higher values for sperm concentration (mean difference 21.7 ± 46.2 times 106 ml‐1, mean ± SD). Motility values showed a corrlation of 0.67 (slope 0.94, P < 0.001) but were estimated lower by the HT than by conventional analysis (mean difference 7.3 ± 21.7%); this was caused by the overestimation of sperm concentration. In the range from 0 to 50% motility the HT yielded lower values and higher values from 50 to 100% motility. Progressive motility values of the HT agreed better with conventional analysis (WHO categories a+b): the mean difference of all values was 0.5 ± 19.5% (r = 0.74, slope = 1.0). The mean lateral head displacement measured by the HT increased with increasing sperm path velocity, but other significant correlations between sperm movement parameters could not be demonstrated. In many instances round cells and debris could not be distinguished from normal sperm. In conclusion, the HT system cannot replace conventional semen analysis in routine diagnosis.
Developmental changes in testicular interstitial cell populations, LH receptors and in the response to hCG in the rat
Authors: LEON MARINA D. , CALVO J. C. , CHARREAU E. H. , CHEMES H. E.
Published: May 2008

High doses of hCG were administered to immature rats of different ages and the animals killed 48 h later. Serum testosterone increased 2 to 4‐fold over control values 48 h after hCG. In‐vitro androgen production showed different patterns according to age. Animals younger than 35 days, when treated with hCG, retained the ability to respond to in‐vitro gonadotrophic stimulation. This ability was lost in testes from rats aged 45 days. The number of free LH‐receptors 48 h after hCG diminished with increasing age to become non‐detectable at 35 and 45 days. In control animals the proportion of differentiated Leydig cells in relation to their mesenchymal precursors increased progressively with age to reach highest values at 45 days. hCG administration induced a shift of the cellular composition of the interstitium toward the more mature cell types. hCG has a predominantly trophic action on mesenchymal precursors in young rats, promoting their differentiation. These effects are minimal in the differentiated Leydig cells in older animals. It is proposed that the observed biochemical responses are the result of the balance between the increase in LH receptors and steroidogenic enzymes in the developing new generation of young Leydig cells and the down‐regulation of receptors and enzymatic lesions in fully differentiated Leydig cells.
In‐vitro production of precursor peptides for thyrotropin‐releasing hormone by human semen
Authors: PEKARY A. E. , REEVE J. R. , SMITH VIERKA P. , FRIEDMAN S.
Published: May 2008

Thyrotrophin‐releasing hormone (TRH) and related peptides occur in high concentrations in human semen. TRH derives from a 242‐amino acid precursor protein, prepro‐TRH, with six repetitive sequences of‐Lys‐Arg‐Gln‐His‐Pro‐Gly‐Lys/Arg)‐Arg‐connected by hydrophobic linking sequences. Antibodies to TRH‐Gly (pGlu‐His‐Pro‐Gly), a final precursor for TRH formation, were used to detect this tetrapeptide as well as other prepro‐TRH fragments which cross‐react with these antibodies. The total TRH‐Gly immunoreactivity decreased significantly after vasectomy. The TRH‐Gly immunoreactivity in semen increased significantly during in‐vitro incubation at 0 or 37d̀C, to a peak value at 5 h, followed by an exponential decline, with t1/2 equal to 11 h at 37d̀C. At 60d̀C, however, the TRH—Gly immunoreactivity rose continuously, attaining, after 20 h, a level 2.2 times that at the start of the incubation (P<0.001). Reversed‐phase high pressure liquid chromatography (HPLC) revealed both hydrophobic and hydro‐philic TRH‐Gly immunoreactive peptides in semen with both classes of peptides increasing significantly with heating to 60d̀C. Cation exchange chromatography of pooled human semen incubated at 60d̀C revealed a 4.3‐fold increase in a TRH‐Gly immunoreactive peak which co‐eluted with synthetic TRH‐Gly, and a 30% increase in another TRH‐Gly immunoreactive peak identified as Glu‐His‐Pro‐Gly. A minor, TRH‐Gly immunoreactive peak increased 50‐fold (P<0.001) during 20 h at 60d̀C. This material co‐eluted with Arg‐Gln‐His‐Pro‐Gly which is formed by enzymic cleavage of the paired basic residues flanking this sequence in prepro‐TRH. When synthetic Arg‐Gln‐His‐Pro‐Gly was incubated with fresh semen at 60d̀C a rapid conversion of most of this peptide to Glu‐His‐Pro‐Gly, Gln‐His‐Pro‐Gly and TRH‐Gly occurred within 30 min. These data are consistent with thermal inactivation of the amidation and degrading enzymes at 60d̀C while the trypsin‐like enzymes which cleave the precursor peptide at the paired basic residues remain relatively unaffected. Because other investigators have found the C‐terminal amidating enzymes to be associated with secretory vesicles and to be co‐secreted with the vesicular contents, we suggest that secretory epithelia of the male reproductive system secrete TRH and TRH‐related precursor peptides along with the alpha‐amidating enzymes which continue processing of prepro‐TRH in the post‐ejaculatory seminal fluid.
Extreme asthenozoospermia and chronic respiratory disease: a new variant of the immotile cilia syndrome
Authors: CHEMES H. E. , MORERO J. L. , LAVIERI J. C.
Published: May 2008

Two patients suspected of suffering from ciliary dyskinesis were investigated. They consulted for primary infertility and chronic respiratory disease. Functional lung studies showed obstructive changes in one patient. Both had immotile sperm with short, thick and rigid tails. Ultrastructural studies of nasal biopsies showed abnormal cilia with almost complete lack of inner dynein arms (mean number of inner arms per axoneme 0.67 ± 1.21 in patient 1 and 1.49 ± 1.17 in patient 2, compared with normal values of 5.3 ± 0.13). Other abnormalities included lack of parallel orientation of cilia and central translocation of microtubular doublets. Electron microscopy of sperm revealed hyperplasia of the fibrous sheath and axonemal disruption. This is the first report of an association of different anomalies in cilia and flagella leading to clinical manifestation of the immotile cilia syndrome. These findings emphasize the need for ultrastructural examination of respiratory cilia in men suffering from fibrous sheath alterations of sperm which so far have not been described in patients with the classical form of immotile cilia syndrome.
Influence of seminal vesicular fluid on the zinc content of human sperm chromatin
Authors: BJORNDAHL L. , KVIST U.
Published: May 2008

Chromatin zinc was studied using X‐ray microanalysis of spermatozoa obtained from split‐ejaculate fractions. Chromatin zinc, expressed as intensity ratio between zinc and sulphur (Zn/S), was unrelated to seminal zinc concentration, but was related inversely to markers of seminal vesicular secretion (fructose concentration and the proportion of zinc bound to ligands of seminal vesicular origin). It is concluded that the content of zinc in sperm chromatin can be reduced by the action of zinc ligands of seminal vesicular origin. An abnormally high contribution of seminal vesicular fluid to sperm‐rich fractions of the ejaculate thus creates a risk of depleting chromatin zinc and thereby impairing zinc‐dependent chromatin stability.
Acidification of epididymal fluid in the boar
Authors: RODRIGUEZ‐MARTINEZ H. , EKSTEDT E. , EINARSSON S.
Published: May 2008

The present study describes the measurement of pH made in vivo in the rete testis fluid and in different regions of the boar epididymis. Furthermore, samples of whole ejaculates, semen fractions, testicular (ductuli efferentes/rete testis), epididymal and deferential fluids collected from the same fertile boars, were analysed for their acid/base status with an automatic blood gas analyser. A pH gradient of acidity was found between the fluid entering the ductus epididymis (pH 7.2) and the region of sperm storage at the cauda (pH 6.5). A significantly lower concentration of bicarbonate ion was found in the cauda epididymidis (3–4 mM) compared to rete testis fluid (30 mM), which might be related to the quiescence of the spermatozoa. A significant increase in extracellular pH and bicarbonate concentration occurred at ejaculation, the bicarbonate levels being 9–10‐fold higher in the semen fraction rich in seminal vesicle fluid, where sperm showed higher motility, than in the cauda epididymis.
Seminal fluid from men with agenesis of the Wolffian ducts: zinc‐binding properties and effects on sperm chromatin stability
Authors: KVIST U. , KJELLBERG S. , BJÖRNDAHL L. , SOUFIR J.‐C. , ARVER S.
Published: May 2008

Zinc‐binding properties were studied in ‘prostatic fluid’, i.e. in seminal plasma from patients with agenesis of the Wolffian ducts, and in split‐ejaculate fractions dominated by seminal vesicular fluid. The effect of seminal fluid, with different zinc‐binding properties, on the stability of zinc‐dependent sperm chromatin was assessed by exposing sperm to 1% sodium dodecyl sulphate (SDS) for 60 min. Citrate was the only zinc ligand in ‘prostatic fluid’, as revealed by gel chromatogra‐phy. Zinc in this fluid enhanced the stability of sperm chromatin. In contrast, the stability of sperm chromatin was decreased in seminal plasma dominated by vesicular fluid. These results are in accordance with the concept that prostatic fluid ensures the appropriate zinc content and stability of sperm chromatin, whereas abundance of vesicular fluid may jeopardize chromatin stability by reducing chromatin zinc content.
Changes in human sperm chromatin stability during preparation for in‐vitro fertilization
Authors: ROSENBORG L. , RAO K. M. , BJÖRNDAHLt L. , KVIST U. , POUSETTE A. , ÁKERLOF EVA , FREDRICSSON B.
Published: May 2008

This study was designed to define the effects of sperm preparation on sperm chromatin stability in relation to in‐vitro fertilization (IVF). Semen samples used for IVF‐embryo transfer (ET) in the treatment of infertility due to tubal factors were studied. Cases with semen variables below reference limits in previous samples were excluded. Sperm were prepared by a swim‐up technique employing either of two different tissue culture media, Ham's F‐10 or Earle's balanced salt solution. Sperm chromatin stability was tested by exposure both to sodium dodecyl sulphate (SDS) only and SDS together with a zinc‐chelating agent, disodium ethylene diamine tetraacetate (SDS‐EDTA). Sperm head swell scores were defined under different experimental conditions and the relationship to sperm motility, morphology, fertilization rate and pregnancy occurrence was tested. No differences were seen between the chromatin stability of sperm from the original sample and that after swim‐up preparation, neither immediately after completion of the swim‐up procedure, nor at the time of insemination of ova. With time, the chromatin became more stable, which occurred to a similar extent both in the original sample and in swim‐up preparations using Ham's F‐10. Otherwise, sperm chromatin stability was unaffected by either of the two media used for swim‐up. At higher incubation temperatures, decondensation in SDS was enhanced. Altogether, no correlation was found between sperm chromatin stability or enhancement of decondensation by temperature and the success of IVF treatment expressed in fertilization rates or pregnancies. The results are reassuring in that only small changes in sperm chromatin stability occurred during the preparation for IVF. As long as semen of presumably good quality is used, these changes in chromatin stability do not seem to be of clinical importance.
Plasminogen activator is involved in the hCG‐induced neutrophil extravasation and vasopermeability increase in the rat testis
Authors: LOUKUSA ANNA‐KAISA , VEIJOLA M. , RAJANIEMI H.
Published: May 2008

The role of proteolytic enzymes in the hCG‐induced increase in testicular vasopermeability and neutrophil extravasation was studied using protease inhibitors. An intra‐testicular injection of hCG together with incubation medium conditioned by polymorphonuclear leucocytes (PMNs) caused a significant increase in vasopermeability and a coincident extravasation of PMN's from the postcapillary venules in the rat testis. When p‐aminobenzamidine, a serine protease inhibitor which inhibits urokinase‐type plasminogen activator, was administered together with hCG in the incubation medium, both the permeability increase and PMN extravasation were prevented. Aprotinin, another serine protease inhibitor, and Eglin C, a specific neutrophil elastase and cathepsin G inhibitor were, however, without effect. None of these inhibitors caused any non‐specific vascular effects in the testis at the concentrations used. These results support the concept that the hCG‐induced increase in vasopermeability in the rat testis is related to extravasation of PMNs and suggest that urokinase‐type plasminogen activator is involved in migration of these cells through the postcapillary venular walls.
Cellular oncogenes in human teratocarcinoma cell lines
Authors: TESCH H. , FÜRBAβ R. , CASPER J. , LYONS J. , BARTRAM C. R. , SCHMOLL H. J. , BRONSON D. L.
Published: May 2008

We have analysed, by Northern blots, the expression of 14 cellular oncogenes in nine cell lines established from human teratocarcinomas. All lines expressed considerable amounts of p53, c‐Ki‐ras2, c‐Ha‐ras1, craf1, N‐myc, and c‐fos. Low level expression of c‐myc was detected in some lines. Southern blot experiments revealed no amplification or rearrangement of the c‐Ki‐ras2, N‐myc or c‐fos genes. Using a rapid dot‐blot screening procedure, based on a combination of in‐vitro amplification of ras‐specific sequences and oligonucleotide hybridization, we could detect no activation of Ha‐ras or Ki‐ras or any unexpressed N‐ras sequences secondary to a point mutation at codons 12, 13, or 61.
Are eosinophil leucocytes involved in the oestrogenic response of the postnatal rat epididymis?
Authors: GAYTÁN F. , ACEITERO J. , BELLIDO C. , PINILLA L. , AGUILAR R. , AGUILAR E.
Published: May 2008

The effects of oestrogens and androgens, alone or in combination, on several epididymal parameters have been studied in 15–day‐old rats after neonatal treatment. Oestrogens induced several responses, such as increased growth of the fibromuscular stroma and eosinophil leucocyte accumulation, whereas the prolif‐erative activity of the epithelium was decreased significantly. Otherwise, the density of intra‐epithelial leucocytes was not modified. Different oestrogen‐induced responses, such as the increase in volume of the fibromuscular stroma and eosinophil leucocyte accumulation were inhibited by treatment with testosterone, whereas dihydrotestosterone had no appreciable effect. This study raises the possibility that eosinophils are mediators of some of the oestrogenic responses in the early postnatal rat epididymis.
Edward Martin (1859–1938). The founding father of modern clinical andrology
Authors: JEQUIER ANNE M.
Published: May 2008

Edward Martin (1858–1938) was the first surgeon to treat successfully the problem of obstructive azoospermia, a feat that he first described in 1902. He subsequently carried out a total of 11 such operations that resulted in seven patencies and three pregnancies. He also demonstrated that the epididymis was the major site of these obstructive lesions. He pointed out the need for semen analysis in diagnosis and also demonstrated that azoospermia could have two causes, namely spermatogenic failure and ductal obstruction. He also made some important observations on sperm movement. He was probably the first clinician to demonstrate surgically ejaculatory duct obstruction, vasal atrophy and congenital absence of the vas. Most important of all, he demonstrated the need for accurate diagnosis in the treatment of male infertility. An argument is put forward that Edward Martin deserves the title of the ‘Founding Father of Modern Clinical Andrology’.
Quantitative assessment of the serum‐induced stimulation of human sperm motility
Authors: de LAMIRANDE EVE , GAGNON C.
Published: May 2008

In previous studies, sperm populations with high motility and fertilizing potential have been isolated using Percoll gradients and/or addition of serum or caffeine, but no objective measurements of the motility parameters affected have been presented. In addition, the types of sperm populations and the incidence of positive or negative responses to these treatments have never been documented. In this paper, the proportion of samples responding positively or negatively to these treatments and the type of motility stimulation observed was measured using the Cell Soft computer‐assisted digital image analysis system. Semen, obtained from the Infertility Clinic, was washed on discontinuous Percoll gradients. Sperm motility parameters were measured in the original semen and after washing through a Percoll gradient in the presence or absence of serum or caffeine. Percoll washing permitted a good recovery of essentially pure sperm. The Percoll treatment had little effect on the percentage of motile sperm but caused a decrease in the linearity‐related parameters. A high proportion of the washed sperm samples supplemented with serum showed an increase in at least one of the motility parameters when compared to sperm populations in the original semen (68% of cases) or after Percoll washing (96% of cases). These effects were maintained for several hours. The stimulation of motility was concentration‐dependent, with an optimum around 25–30% serum. Caffeine (4 mM) increased linear velocity but lowered linearity. In contrast to caffeine, which stimulated sperm motility for less than 1 h, the effects of human serum lasted for more than 16 h.
The physiological significance of pulsatile LHRH secretion in man: gonadotrophin responses to physiological doses of pulsatile versus continuous LHRH administration
Authors: MATSUMOTO A. M. , GROSS K. M. , BREMNER W. J.
Published: May 2008

This study tested whether pulsatile LHRH stimulation of the pituitary is required for normal gonadotrophin secretion in man. Four men with idiopathic hypogonadotrophic hypogonadism (IHH) and presumed endogenous LHRH deficiency were taken off all hormonal replacement for 5–6 weeks, then 5 μg LHRH was administered every 2 h for 1 week in order to prime pituitary gonadotrophin responsiveness. A physiological dose of LHRH (10 μg every 2 h) was then administered in both pulsatile and continuous regimens, in varying order, to each man. Pulsatile LHRH was capable of stimulating LH (as measured by bioassay) and FSH secretion, while continuous administration of LHRH was not. Serum LH, measured by RIA and bioassay, and FSH and free a‐subunit levels, measured by RIA, increased significantly (P<0.05) over pretreatment levels during pulsatile LHRH administration. In contrast, bioactive LH and immunoactive FSH did not change significantly compared to pretreatment values during continuous infusion of the same total LHRH dose, although immunoactive LH and free α‐subunit levels did increase significantly (P<0.05). The ratio of LH bioactivity to immunoactivity was significantly lower during the continuous compared to pulsatile LHRH regimen (P<0.001). Similar serum LHRH levels were achieved during pulsatile and continuous infusions. Serum testosterone and oestradiol levels did not increase significantly from pretreatment levels during either regimen of LHRH administration. It is concluded that a pulsatile LHRH signal pattern is essential for normal pituitary gonadotrophin secretion in men with IHH. Continuous infusion of a physiological dose of LHRH, which produced serum LHRH levels which were indistinguishable from those found during pulsatile administration. failed to stimulate FSH or bioactive LH secretion.
Arginine‐vasopressin induced changes in testicular blood flow
Authors: WIDMARK A. , DAMBER J.‐E. , BERGH A.
Published: May 2008

As arginine‐vasopressin (AVP) has been suggested as a possible paracrine regulator in the testis, the present study was designed to evaluate the effects of AVP on testicular blood flow and vascular permeability. Intratesticular injection of AVP caused a dose‐dependent decrease in total testicular blood flow, with 1 ng AVP being the lowest dose that induced a decrease. The AVP‐induced decrease in blood flow was blocked completely by treatment with an AVP antagonist, but the latter had no effect on its own. High doses of AVP (100 and 1000 ng) decreased testicular IF volume and slightly increased the IF testosterone concentration. At 10 min and 1 h after intratesticular injection of 100 ng AVP testicular blood flow was decreased, followed by an increase at 6 h; testosterone concentrations were increased at both 10 min and 6 h after this treatment. The microcirculatory blood flow pattern (vasomotion) was not influenced by AVP. The present study shows that AVP in low doses influences testicular blood flow rather selectively because AVP was without any major effect on vasomotion, interstitial fluid volume and testosterone production. AVP might therefore be of importance in the regulation of testicular blood flow.
Effect of cryptorchidism on testicular and Leydig cell androgen production in the mouse
Authors: MURPHY LYN , O'SHAUGHNESSY P. J.
Published: May 2008

Unilateral cryptorchidism was induced surgically in adult mice and the effects on testicular and Leydig cell steroidogenesis were studied after 7 weeks. There was a 60% reduction in weight of the cryptorchid testis and this was associated with a significant reduction in intratesticular androgen content, both under basal conditions and following an injection of hCG. Testicular androgen production in vitro was also significantly lower in the cryptorchid testis compared to the scrotal testis, again under both basal conditions (29 ± 6% of control) and in the presence of hCG (46 ± 9% of control). Scrotal testes from the unilaterally cryptorchid animals did not show any significant difference in steroidogenic capacity compared to testes from untreated control animals. The decrease in steroidogenic capacity of the cryptorchid testis was due, at least in part, to a reduction in activity for each Leydig cell. In four experiments, androgen production by Leydig cells isolated from cryptorchid testes was 48 ± 9% of cells from scrotal testes in the presence of a saturating dose of hCG. Under basal conditions the effect was more variable between experiments with steroid secretion by Leydig cells from cryptorchid testes being 58 ± 32% of that for cells from scrotal testes. Leydig cell steroidogenesis in the scrotal testes of unilaterally cryptorchid animals did not differ significantly from untreated controls. These results show that induced cryptorchidism in the mouse causes a significant reduction in Leydig cell activity. This is apparently different from the effects of this procedure on the rat and raises the possibility that intratesticular regulation differs between the two species.
Sperm count and motility influence the results of human fertilization in vitro
Authors: ÅKERLÖF EVA , FREDRICSSON B. , GUSTAFSON O. , LUNELL N.O. , NYLUND L. , ROSENBORG L. , SLOTTE H. , POUSETTE Å.
Published: May 2008

In order to select sperm characteristics that can predict the outcome of in‐vitro fertilization‐embryo transfer (IVF‐ET), semen samples delivered in conjunction with this treatment were studied carefully. We have analysed these data retrospectively in relation to the outcome of treatment. Ninety‐one couples were treated for tubal infertility by IVF‐ET. Fifteen women became pregnant. Sperm were isolated from semen using a swim‐up technique and in most cases 40–80 × 103 (range 20–120 × 103) motile sperm per ovum were used for insemination. The couples were divided into three groups: group A who achieved pregnancies, group B who achieved cleaved ova but no pregnancies, and group C who achieved no ova that were cleaved 48 h after oocyte recovery. Comparisons between these groups showed that some characteristics of the native semen samples and the swim‐up preparations were significantly different: the sperm concentration (P=0.001) and total sperm count (P=0.003) in the native sample, the number of sperm recovered during 30 min of swim‐up (P=0.001), and the specific progressive motility of sperm in the swim‐up preparation, both at the time of insemination and on each day, up to 5 days thereafter (P=0.002–0.028). No pregnancy was achieved with a sperm concentration below 26 × 106 ml‐1 in the native sample. Some of the sperm characteristics studied in this paper may be of value in the pretreatment evaluation for IVF treatment.
Origin of the human seminal plasma motility inhibitor within the reproductive tract
Authors: LUTERMAN M. , IWAMOTO T. , GAGNON C.
Published: May 2008

The origin of a human seminal‐plasma sperm‐motility inhibitor was investigated. Seminal vesicle and prostatic fluids, testes, epididymides, prostate and seminal vesicles were collected from young cadavers. The motility inhibitor, as measured by its capacity to block the motility of demembranated‐reactivated sperm was present in significant amounts only in seminal vesicle fluid. The level of biological activity in this fluid was 13.6‐fold higher than in seminal plasma. However, the concentration of motility inhibitor, as measured by ELISA, using an antibody generated against the motility inhibitor purified from human seminal plasma, was only 1.54‐fold higher than in seminal plasma. The data suggest that the seminal plasma motility inhibitor originates from seminal vesicle fluid and that it is processed into a nine‐fold less biologically active form when seminal vesicle fluid is mixed with other accessory gland secretions at ejaculation.
Testicular steroidogenesis in X/X sex‐reversed mice
Authors: O'SHAUGHNESSY P. J. , ABBOTT D. H. , LEIGH A. J. , CATTANACH B. M.
Published: May 2008

The sex‐reversed X/X Sxr mouse is phenotypically male but lacks germ cells. This provides the opportunity to examine Leydig cell function in the absence of a normal germinal epithelium and without experimental manipulation of the testis. Serum testosterone was lower in Sxr males compared to normal (X/Y) males but there was no significant difference in intratesticular testosterone levels. Serum immunoactive and bioactive luteinizing hormone levels were not significantly different between the two groups. Injection of human chorionic gonadotrophin (hCG) increased intratesticular testosterone in Sxr males more than in normal males although there was no difference in serum testosterone levels. These differences in circulating and intratesticular testosterone levels may be related to reduced blood flow through the Sxr testis. Both basal and hCG‐stimulated androgen production by whole testes in vitro were not significantly different between normal and Sxr males. Androgen production per Leydig cell, however, was significantly reduced in cells from Sxr males; this difference was apparent under basal conditions and following stimulation with hCG, dibutyryl cyclic AMP, 22R‐hydroxycholesterol or pregnenolone. Results show that in the absence of a normal germinal epithelium there is a decrease in the steroidogenic capacity of the Leydig cells although steroidogenesis by the whole testis is not impaired significantly.
Prolactin and α‐1, 4‐glucosidase activity in normal and poorly coagulated human semen
Authors: MANDAL A. , BATABYAL S. K. , BHATTACHARYYA A. K.
Published: May 2008

Prolactin and α‐1,4‐glucosidase levels in seminal plasma were measured in poorly coagulated (I), deficiently coagulated (II) and normally coagulated (III and IV) human ejaculates having 0–20%, 21–50% and 51–100% coagulum respectively 4 min after emission. The prolactin concentration (ng ml‐1, mean ± SEM) in poorly coagulated (5.2 ± 0.48) and deficiently coagulated (7.6 ± 0.72) samples was significantly lower than in the normally coagulated groups III (51–75% coagulum, 8.2 ± 0.43) and IV (76–100% coagulum, 9.9 ± 0.59) as well as the presumably fertile samples (9.2 ± 0.74). A highly significant positive correlation was observed between the profactin level and the percentage coagulum of the ejaculates (r = 0.686, n=58, P<0.001). In contrast, the epididymal marker, α‐glucosidase showed no relationship to seminal coagulation.
Ejaculatory sequence in men with low sperm chromatin‐zinc
Authors: BJÖRNDAHL L. , KJELLBERG S. , KVIST U.
Published: May 2008

The composition of seminal plasma in the sperm‐rich split ejaculate fraction was studied in a group of men with a low zinc content in their sperm chromatin, to evaluate the availability of zinc at ejaculation. Men with low‐chromatin zinc had, in the sperm‐rich split‐ejaculate fraction, high amounts of seminal‐vesicular fluid, a low zinc:fructose molar ratio, and a high percentage of zinc bound to high molecular weight ligands of seminal vesicular origin (HMW‐Zn). This indicates premature admixture of vesicular fluid at ejaculation. It is suggested that the zinc:fructose molar ratio and HMW‐Zn in the sperm‐rich fractions could be used as a measure of the availability of zinc in seminal plasma.