bluuebunny/crossref_metadata_2025 huggingface dataset:
Chromatin alterations induced by freeze‐thawing influence the fertilizing ability of human sperm
Authors: ROYERE D. , HAMAMAH S. , NICOLLE J. C. , LANSAC J.
Published: May 2008

A previous cytochemical study revealed that chromatin alterations could be induced in frozen‐thawed human sperm. In order to evaluate the biological consequences of such chromatin alterations, we evaluated the relationship between these chromatin alterations and the drastic decrease or loss of sperm fertilizing ability after freeze‐thawing. Using acridine orange staining and Feulgen‐DNA quantitative microspectrophotometry, the nuclear variables of sperm were compared using samples which had either recovered, or lost their fertilizing ability after freezing ‐thawing during artificial inseminations, despite good post‐thaw motility. There was a clear decrease in Feulgen‐DNA content and nuclear surface values in sperm which exhibited a loss of fertilizing ability. Thus freeze‐thawing may alter the DNA/nuclear protein relationships and impair the fertilizing ability of human sperm.
Total ligation of the left renal vein in the dog: an inappropriate model for varicocele
Authors: LAVEN J. S. E. , WENSING C. J. G. , MEIJER J.C. , WOLVEKAMP W. T. C. , TEERDS K. J.
Published: May 2008

Induction of varicocele was attempted by ligation of the left renal vein (LRV) in male dogs (Group I). Before the operation and in the 4‐month post‐operative period, sperm count, sperm motility, and sperm morphology of Group I (n = 8) dogs were compared to sham operated animals (Group 11, n = 5). Furthermore, haemodynamics as well as testicular and vascular morphology were studied.

In Group I, changes in diameter and consistency of the spermatic cord were temporary. Semen quality was reduced significantly during the second month after ligation of the LRV, but improved thereafter. Haemodynamic studies revealed that LRV blood pressure was increased significantly in Group I dogs. An extensive venous collateral network replaced the occluded LRV. Retrograde blood flow in the left testicular vein (LTV) was observed only in the proximal part of the LTV of Group I dogs. In Group II dogs numerous pairs of sufficient valves prevented reflux into the LTV. Histological examination revealed that spermatogenesis was not impaired and that the left pampiniform plexus had not changed. The number of Leydig cells was decreased slightly in Group I dogs.

Sufficient valves in the LTV prevented formation of a permanent varicocele.
Inhibition of Sα‐reductase does not simulate the effects of androgen deprivation on porphyrin metabolism by Harderian glands from the male Syrian hamster
Authors: MENENDEZ‐PELAEZ A. , BUZZELL G. R. , REITER R. J.
Published: May 2008

As Sα‐reductase is involved in the metabolism of indolamines in the Harderian glands of Syrian hamsters, we have compared the effects of androgen deprivation and the administration of the potent Sα‐reductase inhibitor N, N‐diethyl‐4‐methoxy‐3‐oxo‐4‐aza‐5‐androstene‐17‐carboxamida on Harderian porphyrin metabolism. Ten days after castration, porphyrin levels had increased to 40 times the values of intact hamsters. However, the inhibition of Sa‐reductase, which resulted in a significant decrease in the weight of accessory sex glands, did not modify porphyrin concentrations within the Harderian glands. It is concluded that, contrary to the situation for indolamine metabolism, testosterone per se is the main androgen involved in the sexual differences observed in Harderian porphyrin metabolism.
Non‐specific uptake of IgG by rat epididymal tubules in vitro
Authors: YEUNG C. H. , BERGMANN M. , COOPER T. G.
Published: May 2008

Tubules isolated from the initial segment, caput and corpus regions of the rat epididymis were incubated for 4 h in medium containing rat IgG or bovine‐serum‐albumin (10 mg ml‐1) and processed for light microscopy immunocytochemistry using gold‐labelled anti‐rat IgG and silver staining or peroxidase/anti‐peroxidase techniques. Distribution of IgG was confined to the peritubular matrix only, with no detectable uptake into the epithelium or lumen. In tubules incubated with rat IgG coupled to gold particles as tracer and fixed for electron microscopy, the IgG‐gold complex failed to penetrate to the base of the epithelium. When IgG‐peroxidase conjugates were used, sparse enzyme activity was localized in small basal and apical vesicles and multivesicular bodies of the principal cells. However, this uptake could not be inhibited by the presence of a 30‐fold excess of non‐labelled IgG, and the same distribution of enzyme activity was also observed when tubules were incubated with a similar activity of horseradish peroxidase alone. These findings indicate that there is no detectable receptor‐mediated transport of IgG across the epididymal epithelium, although non‐selective transfer could occur via fluid‐phase endocytosis.
Induction of the acrosome reaction in sperm by exposure to low temperature increases their rate of fusion with zona‐free hamster oocytes
Authors: Sanchez R. , Schill W.‐B.
Published: May 2008

The fusion rate of human sperm with zona‐free hamster ova was investigated after induction of the acrosome reaction by exposure to a low temperature (4°C). Sperm were collected from 14 patients, and selected by the ‘swim‐up’ method. The sperm were incubated for 24 h at either room temperature (control group) or at 4°C (low temperature group), followed by additional incubation at 37°C for 3 h. The mean sperm penetration rate, number of swollen sperm heads as well as the number of sperm attached to the oocyte increased significantly after exposing sperm to low temperature. The sperm penetration rate showed a significant correlation (Spearman test, r= 0.572, n= 28, P <0.0035) with the acrosome reaction in the low temperature group. These results were associated with an increase in the rate of penetration of hamster ova observed in this study, presumably due to the increase in induction of the acrosome reaction by low temperature. Incubation of sperm at low temperature might be useful in the evaluation of so‐called false negative results in the zona‐free hamster test.
Capacitation and the acrosome reaction in sperm from men with various semen profiles montored by a chlortetracycline flrorescence assay
Authors: KHOLKUTE S. D. , MEHERJI P. , PURI C. P.
Published: May 2008

Sperm obtained from groups of men with various semen profiles were incubated for 8 h in BWW medium containing human serum albumin to promote capacitation. Capacitation and the acrosome reaction were monitored by a chlortetracycline (CTC) fluorescence assay. Four distinct CTC patterns were observed on the sperm head. No significant difference was observed in the time‐course curve of these CTC patterns in sperm obtained from normozoospermic, asthenozoospermic and oligo‐zoospermic men. Spontaneous and A23187‐induced acrosome reactions were also comparable in these groups. However, in sperm obtained from teratozoospermic and polyzoospermic men, the increase in CTC pattern associated with capacitation appeared slower and sluggish. In these two groups, the induced acrosome reaction was also significantly lower when compared to that in the other three groups of men. In polyzoospermia, the spontaneous acrosome reaction was significantly lower when compared to all the other groups. Fresh sperm would not undergo the acrosome reaction following A23187 treatment. The results of this study indicate sluggish (defective) capacitation and inability of capacitated sperm to undergo induced acrosome reaction in teratozoospermic and polyzoospermic men as evaluated by the CTC method.
Testosterone increases serum 1,25–dihydroxyvitamin D and insulin‐like growth factor‐I in hypogonadal men
Authors: HAGENFELDT Y. , LINDE K. , SJÖBERG H.‐E. , ZUMKELLER W. , ARVER S.
Published: May 2008

The short‐term metabolic effects of testosterone treatment on circulating levels of 1,25–dihydroxyvitamin D and insulin‐like growth factor‐I (IGF‐I) were studied in 13 hypogonadal men. The study group included 11 men with Klinefelter's syndrome, with varying degree of androgen deficiency, and two men with secondary hypogonadism.

Pretreatment levels of 1,25–dihydroxyvitamin D, vitamin D‐binding protein and IGF‐binding protein‐I were all within the normal range. The levels of IGF‐I were lower than normal in 5/11 of the Klinefelter patients and in one patient with GH‐deficiency. Testosterone treatment increased circulating total 1,25–dihydroxyvitamin D significantly from 75 ± 4 pmol l‐1 (mean ± SEM) to 86 ± 4 (P<0.01) and the free 1,25–dihydroxyvitamin D‐index from 1.95 ± 0.11 to 2.39 ± 0.12 (P<0.01). Serum levels of IGF‐I increased from 117 ± 22 ug/l to 143 ± 23 (P<0.01) during androgen treatment. No significant effects on levels of IGF‐binding protein‐I were seen.

It is concluded that androgen therapy increases the availability of 1,25–dihydroxyvitamin D and the level of IGF‐I, which may be important links in the action of testosterone.
Testosterone production in mature scorbutic mutant rats unable to synthesize ascorbic acid
Authors: ITO Y. , TSUJI M. , TERADA N. , MIYANO M. , MORI H.
Published: May 2008

The effect of a deficiency of ascorbic acid (AsA) on in‐vivo testosterone production in mature male rats was investigated using a mutant strain of rats (ODS rats) unable to synthesize AsA. Male 60–day‐old rats were fed AsA‐deficient lab chow for 28 days with [ODS(+)] or without [ODS(‐)] AsA supply. The AsA levels in the liver of ODS(‐) rats were undetectable and those in the testes decreased to about 25% of those in ODS(+) rats by day 28. Plasma LH levels in ODS(‐) rats decreased to about 30% of those in ODS(+) rats by day 28. However, there were no significant differences in the plasma levels of testosterone, or in the relative weights of seminal vesicles and ventral prostates between ODS(+) and ODS(‐) rats. Plasma levels of testosterone in ODS(‐) rats after a single injection of 200 IU hCG changed similarly to that in ODS(+) rats. The metabolic clearance rate of testosterone was also the same at 60 min after an intravenous injection of [3H]testosterone in both groups. These results indicate that AsA‐deficiency in adult rats causes no significant change in basal plasma levels of testosterone or in the response to hCG, despite decreased plasma LH levels.
Deficiency in major high molecular‐weight seminal proteins in men producing poorly coagulating and alkaline liquid ejaculates
Authors: MANDAL A. , BHATTACHARYYA A. K.
Published: May 2008

Using SDS‐PAGE three major high molecular‐weight coagulum proteins (57, 75 and 79 kD) were identified in unliquefied human ejaculates. Washed coagulum was prepared at pH 4.5 and demonstrated negligible proteolysis as determined by a highly sensitive fluorescamine‐based method. Laser densitometric scanning of these high molecular‐weight proteins in unliquefied ejaculates in relation to their coagulational failure revealed that the total content of 57, 75 and 79 kD proteins in whole semen represented ˜ 44% of levels in the normally coagulated, ˜ 24% in poorly coagulated (P<0.005) and ˜ 3% in the non‐coagulated (alkaline liquid, P<0.001) samples. The findings imply that these predominant high molecular‐weight proteins play a major role in human seminal coagulum formation.
Sperm nuclear instability and staining with aniline blue: abnormal persistance of histones in spermatozoa in infertile men
Authors: FORESTA C. , ZORZI M. , ROSSATO M. , VAROTTO A.
Published: May 2008

During mammalian spermiogenesis, replacement of the somatic histones by basic proteins, the protamines, allows normal sperm nuclear condensation. In this study we have evaluated the degree of chromatin compaction in spermatozoa from 191 infertile subjects, affected by different testicular disorders, compare with that in 50 fertile sperm donors (controls). In infertile men, there was a higher percentage of unstable spermatozoa after incubation with sodium dodecyl sulphate (SDS) and of stained spermatozoa after staining with aniline blue (P <0.001 vs. controls). Furthermore, a positive linear correlation was found between SDS‐unstable spermatozoa and stained spermatozoa (P <0.001), suggesting that sperm instability was related to a defect in histone‐replacement by sperm‐specific nucleoproteins, protamines. When the patients were considered according to pathology, high sperm nuclear instability and a high percentage of stained spermatozoa were detected in groups affected by varicocele, idiopathic infertility and in patients with a history of unilateral cryptorchidism. In the latter group the same alterations were observed even when the cryptorchid testis had been removed during surgery. In the group with a past history of mumps orchitis these parameters did not show any difference when compared with controls.
Semen analysis and fertility prognosis in andrological patients
Authors: HOLLAND‐MORITZ H. , KRAUSE W.
Published: May 2008

Semen analyses of 529 men who consulted our department due to infertility problems, were related to the time period prior to conception, with factors adversely affecting the fertility of the female partner taken into consideration. The statistical method used was Cox's proportional‐hazard model of regression. Untransformed, logarithmically transformed and dichotomized semen analysis variables were included in the calculations. The relationship between the following parameters and the probability of conception was examined: sperm count, sperm motility, progressive sperm motility, morphology and sperm motility remaining 24 h after ejaculation. All variables co‐varied with the probability of conception; however, the exact type of relationship could not be determined by regression analyses. Cox's model assumes an exponential relationship. Our data suggest that this assumption is not suitable for fertility investigations. Using conventionally defined limiting values for normal and pathological semen quality, statistical analysis yielded significant differences in fertility between both categories for all of the variables considered; in the stepwise regression analysis, however, it could be shown that progressive motility and morphology alone were sufficient to discriminate between normal and pathological semen quality. The results are interpreted as indicating that, as a result of semen analysis, it is possible to predict the individual probability of conception if the exact shape of the relationship can be determined, which, up to now, has not been accomplished.
Residual sperm function in oligozoospermia induced by testosterone enanthate administered as a potential steroid male contraceptive
Authors: WALLACE E. M. , AITKEN R. J. , WU F. C. W.
Published: May 2008

To investigate the fertility of men who remain oligozoospermic despite sex steroid suppression, the in‐vitro fertilizing capacity of residual spermatozoa was assessed in 30 men receiving intramuscular testosterone enanthate (TE). Spermatozoa were prepared by either Percoll or repetitive centrifugation/washing. Although the mean (± SEM) pretreatment zona‐free hamster oocyte penetration (HOP) rates were similar (59.4 ± 10.1 and 63.8 ± 10.8%), following the induction of oligozoospermia the Percoll‐prepared spermatozoa exhibited a penetration rate (26.9 ± 10.2%) which was markedly greater than that obtained for sperm prepared by repetitive washing (0 ± 0%). In addition, the partners of two men exhibiting a HOP test with Percoll‐prepared spermatozoa, conceived despite a sperm concentration of 3 × 106ml‐1 and a negative HOP test with spermatozoa prepared by repetitive washing. These results suggest that Percoll preparation optimizes the assessment of in‐vitro sperm function and that the fertility of men with TE‐induced severe oligozoospermia is suppressed but not abolished.
Infection in the male reproductive tract. Impact, diagnosis and treatment in relation to male infertility
Authors: Purvis K. , Christiansen E.
Published: May 2008

The following are the conclusions that can be derived from a review of the literature regarding the role of infection in the aetiology of male infertility. (i) Temporary inflammatory episodes in the male reproductive tract which are self‐limiting are probably common. (ii) Caution should be exercised in the use of leukospermia or bacteriospermia as parameters for glandular infection. (iii) There is a need for alternative techniques for detecting non‐symptomatic deep pelvic infections in the male; one technique of great promise is rectal ultrasound. (iv) Rectal ultrasound indicates that a large number of men with poor sperm quality have a non‐symptomatic, chronic prostatovesiculitis. (v) Increasing evidence implicates Chlamydia trachomatis as being a major cause of chronic non‐bacterial prostatitis. (vi) An important aspect of chlamydial infections in men may be that the male accessory sex glands may function as reservoirs for the organism, increasing the probability of infection in the female. (vii) Ureaplasma urealyticum may also play an important aetiological role in male infertility but its significance is confounded by its acknowledged function as a commensal in the reproductive tract. (viii) One of the manifestations of male reproductive tract infectiw is the induction of sperm autoantibodies. (ix) There is a need for more systematic controlled studies of the effects of antibiotic treatment on sperm quality with different preparations for extended periods using patient groups in which a glandular infection has been verified, e.g. by rectal ultrasonography.
The time‐response and magnitude of HCG‐induced vascular changes are different in scrotal and abdominal testes
Authors: Hjertkvist M. , Bergh A.
Published: May 2008

Adult unilaterally cryptorchid rats were injected with 50 IU human chorionic gonadotrophin (hCG). At 4, 8, 24 and 72 h after treatment, testicular vascular permeability was studied by injecting colloidal carbon intravenously. The number of blood vessel profiles labelled with carbon was increased by hCG in both types of testes, but the response was more sustained in abdominal than in scrotal testes. The number of polymorphonuclear leucocytes (PMNs) accumulating in testicular blood vessels and migrating into the interstitial space in response to hCG treatment was also measured. The volume density of intravascular and interstitial PMNs was increased in both types of testes but the peak response was larger in scrotal than in abdominal testes. PMN accumulation and vascular leakage were apparently correlated in the scrotal but not in the abdominal testes.

Testicular interstitial fluid (IF) was collected from intact and unilaterally cryptorchid adult rats. The IF was diluted with sterile buffer and injected intracutaneously in test animals. The vascular permeability response was assessed by measuring the leakage of Evans blue into the injection sites. IF from scrotal and abdominal testes increased vascular permeability in the skin. The response was rapid and transient. IF collected from rats given hCG 24 h earlier did not increase vascular permeability. The vascular permeability response to IF was reduced slightly in neutrophil‐depleted animals. The inflammation mediator present in IF cannot explain the kinetics and magnitude of the hCG‐induced changes in vascular premeability in intact or unilaterally cryptorchid rats.
Qualitative assessment of thermal and evaporative adjustments of human scrotal skin in response to heat stress
Authors: CANDAS V. , BECMEUR F. , BOTHOREL B. , HOEFT A.
Published: May 2008

To study scrotal thermoregulation and its efficacy to work against heat accumulation, five subjects were exposed to four experimental conditions under which core and skin temperatures and sweat evaporative responses of various skin surfaces—chest, abdomen and scrotum—were compared. The temperature response of the scrotal area exhibited the largest inertia, and this observation is likely to be the consequence of heat exchange via the vascularization of testes and scrotum which is more efficient than in other parts of the body in limiting local heat storage, thus alleviating heat stress of the testis. The pulsatile nature and the synchronous pattern of the scrotal evaporative heat loss indicate that scrotal sweating takes place, although the gradient response appeared to be less marked than elsewhere in the body. Relatively low and inert scrotal temperature can partly explain this poor local drive for sweating.
Serum levels of testosterone and oestrogens in normal and infertile stallions
Authors: INOUE J. , CERBITO W. A. , OGURI N. , MATSUZAWA T. , SATO K.
Published: May 2008

The serum concentrations of testosterone and oestrogens were determined in stallions classified as geldings, normal (according to age) or infertile (azoospermic). There were significant differences in testosterone and oestrogen levels between the groups. Normal concentrations of testosterone and total oestrogens were attained after 16 months of age. Castrated and immature horses (<15 months of age) had lower levels of testosterone and total oestrogens than did mature and infertile stallions. Azoospermic stallions had lower levels of testosterone and total oestrogens than did normal mature horses except in one case (B4) in which the level of total oestrogens was increased markedly. Critical ellipses for testosterone and total oestrogens were obtained for both groups (6–15‐month‐old colts and stallions aged >4 years). The results suggest that serum levels of testosterone and total oestrogens may provide a sensitive index of the endocrine function of the testis in male horses.
Effect of chronic alcoholism on semen—studies on lipid profiles
Authors: GOMATHI C. , BALASUBRAMANIAN K. , BHANU N. VIJAYA , SRIKANTH V. , GOVINDARAJULU P.
Published: May 2008

The effect of chronic alcoholism on various seminal parameters (sperm concentration, rate of forward motility, percentage of abnormal spermatozoa, lipid profiles of seminal plasma and spermatozoa) was studied together with the serum levels of testosterone and oestradiol. In chronic alcoholics there was a marked reduction in sperm concentration and in the rate of their forward motility, and increase in the number of spermatozoa with morphological abnormalities when compared to age‐matched normal fertile subjects. Serum levels of testosterone were decreased while oestradiol levels were increased in chronic alcoholic men. Studies of lipid profiles showed a marked decrease in the total phospholipid concentration in spermatozoa, primarily in sphingomyelin, phosphatidyl choline and ethanolamine fractions. The cholesterol phospholipids ratio in spermatozoa was increased in alcoholics. In the seminal plasma of chronic alcoholics, there was a decrease in total lipid, in glyceride glycerol and in free and esterified cholesterol. Of the phospholipid classes, sphingomyelin and phosphatidyl ethanolamine showed a significant reduction. In general, the present study provides evidence for the adverse effects of chronic alcoholism on serum hormones, sperm count, morphology, motility and seminal lipid profiles. These may be responsible for the fertility disorders common in chronic alcoholics.
Reactive oxygen species in semen of infertile patients: levels of superoxide dismutase‐ and catalase‐like activities in seminal plasma and spermatozoa
Authors: ZINI A. , de LAMIRANDE E. , GAGNON C.
Published: May 2008

Reactive oxygen species (ROS) can be detected in the semen of 40% of infertile men, whereas none is detected in semen from normal men. The ROS detected in semen are a reflection of the imbalance between ROS production and degradation. The aim of the present study was to determine whether a lowered scavenging capacity or an increased production of ROS was responsible for the ROS detected in semen samples from infertile men. Two activities were investigated: (1) catalase‐like activity, which is responsible for the degradation of H2O2, and (2) superoxide dismutase‐like (SOD‐like) activity which is responsible for the degradation of O2‐‐. Catalase‐like and SOD‐like activities were found in whole seminal plasma, in dialyzed seminal plasma (> 12 kD), in an ultrafiltrate of seminal plasma (< 5 kD) and in spermatozoa. There was no significant difference in the SOD‐like activities measured in spermatozoa, or in seminal plasma (whole or fractionated) from samples that did or did not produce ROS. SOD‐like activity originated mostly from the high molecular weight components of seminal plasma. However, the catalase‐like activity of whole seminal plasma and of spermatozoa was significantly greater (P = 0.01) in those samples that produced ROS as compared to those that did not. The catalase‐like activity in dialyzed seminal plasma, and an ultrafiltrate of seminal plasma from semen samples that did or did not produce ROS were not statistically different. The catalase‐like activity of the seminal plasma originated equally from high and low molecular weight components. In conclusion, the data suggest that the ROS detected in the semen of infertile patients are likely due to increased ROS production rather than to decreased ROS scavenging capacity.
Is testosterone essential for maintenance of normal morphology in immature rat Leydig cells?
Authors: MISRO M. M. , GANGULY A. , DAS R. P.
Published: May 2008

Selective deprivation of gonadotrophins in prepubertal rats by administration of a GnRH antagonist (Ac‐D2Nal1, D4ClPhe2, DTrp3, DArg6, DA1a10‐GnRH; GnRH code: 103–289–10, National Institutes of Health, USA) for 3 weeks, initiated at 20–22 days of age, induced morphological changes in the Leydig cells, including thickening and indentation of the nuclear margin, pyknosis and elongation of the nuclei. Mean nuclear diameter was reduced to 22% of that in the controls. Under the electron microscope the cells exhibited reduced volume of the nucleus and cytoplasm and the plasma membrane was irregular. This abnormal appearance of the Leydig cells improved marginally in 20–30% of the Leydig cells and their mean nuclear diameter increased to 39% of the control level after FSH supplementation (20 μg ovine FSH/day). Normal morphological integrity of the Leydig cells consisting of round or oval nuclei, a smooth nuclear and cellular margin and the original mean nuclear diameter was restored completely when testosterone (30 μg/day) was administered to GnRH antagonist‐treated rats, with or without simultaneous administration of FSH; in these rats testosterone levels in blood were also restored to normal. These findings indicate that testosterone may be important for the maintenance of normal Leydig cell morphology in the rat.
Alterations in testicular function after endotoxin injection in the boar
Authors: WALLGREN MARGARETA , KINDAHL H. , RODRIGUEZ‐MARTINEZ H.
Published: May 2008

Twelve mature boars were injected intravenously with endotoxin from Salmonella typhimuvium. Blood plasma was analysed for 15‐ketodihydro‐PGF2α, LH and testosterone. The boars were castrated at various times after endotoxin administration and the testes examined by light and electron microscopy. The levels of 15‐ketodihydro‐PGF2, and LH rose immediately after endotoxin injection followed by an increase in testosterone levels, and in five boars a second increase in LH levels was observed. Morphological examination of the testes revealed infiltration of polymorphonuclear neutrophils (PMN) into the testicular interstitium. The Leydig and myoid cells showed morphological changes.

Alterations were also present in the seminiferous epithelium among Sertoli cells, spermatocytes and spermatids. The results indicate that endotoxins can exert a negative effect on testicular function in the boar. Except for the initial increase in LH levels, which is not clearly understood, the hormonal changes are thought to be mediated by alterations in Leydig cell function. The alterations further support previous findings in the boar, and indicate that there is short‐term, moderate damage to the seminiferous epithelium following endotoxin injection.
Influence of three different preparation techniques on the results of human sperm morphology analysis*
Authors: MESCHEDE D. , KECK C. , ZANDER M. , COOPER T. G. , YEUNG C.‐H. , NIESCHLAG E.
Published: May 2008

Using 158 unselected semen samples the present study has analysed how the results of sperm morphology assessment were influenced by different techniques for preparing the slides for microscopic assessment. All three techniques compared, the Papanicolaou stain (PAP), the Shorr stain (SHO) and the ‘wet preparations’ protocol (WET) are currently recommended by the World Health Organization for use in andrology laboratories. Mean percentages of morphologically normal spermatozoa were identical on PAP and SHO slides (31.1%), but were significantly lower in wet preparations (12.3%). Wide divergence of results obtained with the three different methods was also found with respect to the percentage of sperm with head, midpiece and tail defects and two ‘indices of teratozoospermia’. For the majority of parameters assessed, linear regression analysis revealed a poor correlation between evaluations of PAP, SHO and WET slides (r values ranging from 0.01 to 0.87). We conclude that only one standard method should be recommended for the preparation of morphology slides in order to ensure inter‐laboratory comparability of results and to enhance the value of sperm morphology analysis for predicting fertility.
Urine as an alternative to urethral swabs for the diagnosis of Chlamydia trachomatis in infertile males
Authors: RAMUTHAGA T. N. , BORNMAN M. s. , MAHOMED M. F. , BOOMKER D. , GREEF A. S. , CREWE‐BROWN H. H. , REIF S.
Published: May 2008

Swabbing the urethrae of men has been the traditional approach for collecting specimens for detection of Chlamydia trachomatis. Recently, however, urine testing using enzyme immunoassay has yielded promising results. A total of 105 patients attending the Andrology Clinic at Ga Rankuwa Hospital, Medunsa were included in the study. These patients were asymptomatic and had no urethral discharge. Three endo‐urethral swabs and first‐catch urine were collected fiom each patient. The urethral swabs were used for enzyme immunoassay (EIA) (IDEIA 111), tissue culture and direct immonufluorescent antibody (DFA) test (IMAGEN) to detect C. trachomatis. In addition about 15–30 ml of first‐catch urine, or urine collected at least 2h after the previous micturition, was collected for each patient for EIA testing. Fifteen (14.3%) of 105 patients were positive on urethral swab EIA, in comparison with the DFA test in which 14 (13.3%) were positive. Eight (7.8%) were positive in tissue culture. Urine EIA was positive in 17 (16.2%) patients, of whom five (4.8%) were positive in urine EIA only. All EIA positive urines were confirmed by DFA. We recommend that first‐catch urine or urine collected at least 2h after the previous micturition in infertile males may be considered a suitable alternative to urethral swab for chlamydial diagnosis because it is noninvasive and nontraumatic.
Simultaneous measurement of sperm LDH, LDH‐X, CPK activities and ATP content in normospermic and oligozoospermic men
Authors: ORLANDO C. , KRAUSZ C. , FORT G. , CASANO R.
Published: May 2008

Total lactate dehydrogenase (LDH), LDH‐X, creatine phosphokinase (CPK) activities and ATP content were measured in washed spermatozoa from 188 normospermic and 94 oligozoospermic men. These four biochemical parameters were significantly higher (p>0.00l) in spermatozoa from oligozoospermic men than in those from normospermic men and were related inversely to sperm concentration. In oligozoospermic men the activity of sperm LDH and LDH‐X was related sipficantly to sperm CPK activity (r = 0.5486 and r = 0.4442, respectively), whereas this relationship was less evident in normospermic men (r =0.4124 andr= 0.3844, respectively). Sperm ATP content was related weakly to sperm LDH, LDH‐X and CPK activities both in oligozoospermic and normospermic men. After separation on a discontinuous (40% and 80%) Percoll gradient LDH, LDH‐X, CPK activities and ATP content in 80% separated sperm fractions from 48 normospermic and 30 oligozoospermic men were found to be significantly lower than in the initial semen sample in both groups of subjects. The 80% separated sperm fi‐actions fiom oligozoospermic and normospermic men did not differ in their LDH or LDH‐X activities or ATP content, whereas sperm CPK activity was still significantly higher in oligozoospermic men. These data suggest that, of the four parameters studied, high levels of intracellular CPK activity are the most relevant biochemical feature of spermatozoa from oligozoospermic men.
Seminal plasma lead, cadmium and zinc in relation to tobacco consumption
Authors: OLDEREID N. B. , THOMASSEN Y. , PURVIS K.
Published: May 2008

The total quantity of zinc in the ejaculates of smokers was sipficantly lower than in non‐smokers. This was not related to a significant increase in the quantities of seminal cadmium or lead, or to a decrease in sperm quality in the smoking group. It appears that tobacco consumption may have to exceed 20 cigarettedday before a noticeable increase in seminal cadmium can be recorded. It is suggested that this reduction in zinc secretion may jeopardize the content of chromatin zinc, and thereby the stability of the sperm chromatin. This may then contribute to reproductive failure or have consequences for fetal development.
Objective assessment of spermatogenesis in men with functional and anatomic obstruction of the genital tract*
Authors: HIRSCH I. H. , SEDOR J. , KULP D. , McCUE P. J. , STAAS W. E.
Published: May 2008

Experimental rodent models simulating the condition of neurogenic infertility have drawn attention to the role of potential epididymal dysfunction as an underlying cause.

This functional obstruction of the genital tract is comparable to the outcome of genital tract obstruction after vasectomy, and may explain the common finding of asthenospermia in both groups following either stimulated semen recovery or vasovasostomy, respectively. Since spermatogenic dysfbnction has been reported in spinal cord injury, the relative roles of defective sperm production and sperm transport remain to be determined in men with neurogenic infertility. The objective of this study was to compare the levels of spermatogenesis in groups of vasectomized men and those with spinal cord injury, using objective measurement criteria for spermatogenesis.

Groups of 10 spinal cord‐injured and six vasectomized men matched for age and duration of disease, underwent incisional testicular biopsy. The specimens were divided equally for parallel quantitation of spermatogenesis by both quantitative cytometry and DNA flow cytometric analysis. Quantitative parameters showed similar values for both groups with reference to mean tubular wall thickness, mean tubular concentration of spermatids and Sertoli cells, as well as the mean spermatid: Sertoli cell ratio per tubule. Additionally, similar percentages of lN, 2N and 4N cells were found in both groups‐ Based on these preliminary findings this study provides a clinical correlation supporting the experimental observation that both anatomical and functional obstruction of the male genital tract exert a similar although minor spermatogenic insult, and that in both the putative cause for neurogenic infertility is more likely to be at the post‐testicular level.
Affinity sites for N‐acetyl‐β‐D‐glucosaminidase on the surface of rat epididymal spermatozoa*
Authors: BARBIERI M. A. , SOSA M. A. , COUSO R. , IELPI L. , MERELLO S. , TONN C. E. , BERTINI E.
Published: May 2008

The binding of N‐acetyl‐β‐D‐glucosaminidase from rat epidldymal fluid to the surf‐ace of spermatozoa from the cauda epididymis was measured in the presence of sugars, its phosphorylated derivatives, or after treatment of the cells or the enzyme with agents that alter the integrity of proteins or carbohydrates. The binding was saturable, with a K, in the nanomolar range, was inhibited with phosphorylated derivates of fructose, and did not depend on Ca2+ showing that it is different from the mannose 6‐P‐recognizing system existing in other tissues for this and other acid hydrolases. Treatment of the cells with sodium periodate or trypsin inhibited the binding, showing that a glycoprotein of the plasmalemma is involved in the affinity site. Fructose or phosphorylated derivates were not detected in the proteins of the epididymal fluid with HPLC. However, with the method used, the presence of these compounds cannot be ruled out, if among the proteins of the fluid there are only a small number of acid hydrolases containing this sugar.
Relationship between semen characteristics, a‐glucosidase and the capacity of spermatozoa to bind to the human zona pellucida
Authors: ALI H. BEN , GUERIN J. F. , PINATEL M. C. , MATHIEU C. , BOULIEU D. , TRITAR B.
Published: May 2008

Unfertilized oocytes from an in‐vitro fertilization programme were stored in different saline solutions and then utilized in a zona binding assay (ZBA). The four saline solutions tested were identical with regard to the capacity of the zona to bind sperniatozoa provided by healthy donors. Spermatozoa from 150 infertile patients were tested in the ZBA. The number of spermatozoa bound to the zona correlated positively with sperm concentration, the percentage sperm motility and the percentage of morphologically normal spermatozoa. The population was then divided into two groups according to the level of α‐glucosidase activity, an epididymal marker. The average number of spermatozoa bound to the zona was diminished in the group with low α‐glucosidase activity, even when considering strictly equivalent classes of sperm concentration, motility and morphology, respectively.
Modulation of ornithine decarboxylase activity by prolactin in seminal vesicles of the rat
Authors: GONZALEZ S. I. , SUESCUN M. O. , RULLI S. B. , ESTIVARIZ E. , CALANDRA R. S.
Published: May 2008

The stimulatory effect of prolactin on sexual accessory glands is well established, though the mechanism of trophic action remains poorly understood. We have therefore assessed the effect of high levels of prolactin on ornithine decarboxylase activity and polyamine content in seminal vesicles (SV) of adult rats. Hyperprolactinaemia was induced by implantation of tissue fragments of a prolactin‐secreting tumour 7315a, and the rats killed at 35 days post‐inoculation. Serum levels of prolactin were increased significantly (p<0.001) in tumour‐bearing rats. Levels of testosterone in serum were reduced markedly, whereas LH levels remained unchanged. SV weight in the experimental group was not significantly different from that in the corresponding control group. A clear increase in ornithine decarboxylase activity of SV was observed (p<0.001) in the hyperprolactinaemic rats. However, there was no change in the polyamine content of SV in tumour‐bearing rats, compared to the control group. These results indicate that ornithine decarboxylase activity in SV of adult rats is regulated mainly by prolactin. This experimental model may be useful for identification of some of the events involved in the trophic action of prolactin.
The effect of experimental cryptorchidism on the phosphorus NMR spectrum of the rat testis
Authors: VAN DER GROND J. , DIJKSTRA G. , VAN ECHTELD C. J. A.
Published: May 2008

Magnetic resonance (MR) spectroscopy of the cryptorchid rat testis was used to test whether changes in the MR spectra of the rat testis might be a more sensitive indicator of changes in the metabolic status of germ cells in the testis rather than simply the cell types present. Testes of adult Wistar rats before and during 42 days of experimental cryptorchidism were investigated by in‐vivo 31P MR spectroscopy. Results were compared to MR studies of the synchronized developing testis. The testicular phospho‐monoester/ATP (PM/ATP) ratio was dependent only on the cell types present, and showed the same characteristics for each cell type present in the degenerating testis as in the developing testis. The testicular phosphodiester/ATP (PD/ATP) ratio decreased rapidly when the number of round and elongated spermatids was reduced. Similar effects, although less pronounced, were seen in the developing testis. The pH decreased rapidly after cryptorchidism, and was related inversely to the PM/ATP ratio, which was also observed in the developing testis. This study demonstrates that MR spectroscopy monitors the cell types present in the rat testis rather than its metabolic status.
The potential of mild testicular heating as a safe, effective and reversible contraceptive method for men
Authors: MIEUSSET ROGER , B'UJAN LOUIS
Published: May 2008

A preliminary assessment of the contraceptive efficacy of a daily mild increase (1–2°c) in testicular temperature during waking hours is reported in nine couples using two techniques of non‐surgical fixation of the testes close to the inguinal canal. With technique 1, immobilization was achieved by passing the penis and the empty scrotum through a hole made in close‐fitting underwear; there was one pregnancy, hm a man who stopped the heating after 7 weeks, for 42 cycles of exposure in three couples. With technique 2, immobilization was achieved by adding a ring of soft material surrounding the hole in the underwear; there was no pregnancy for 117 cycles of exposure in six couples. Reversibility and safety were assessed. These preliminary results suggest that a daily mild increase in testicular temperature could be a potential contraceptive method for men.
Pentoxifylline acts synergistically with A23187 to increase the penetration of zona‐free hamster oocytes by cryopreserved human spermatozoa
Authors: FORD W. C. L. , REES J. M. , McLAUGHLIN E. A. , LING L. , HULL M. G. R.
Published: May 2008

The number of cryopreserved human spermatozoa which penetrated zona‐free hamster oocytes afier stimulation with 2μmol A23187 per litre was increased by the hrther addition of 0.6 or 3.6 mmol pentoxifjrlline per litre. With spermatozoa prepared by washing by repeated centrifugation, the median numbers of sperm headd/egg were 1.9, 7.9 and 10.8 in the presence of 0, 0.6 or 3.6 mmol pentoxifylline per litre, respectively. A similar effect was observed with spermatozoa prepared on a Percoll gradient. As A23187 inhibited sperm motility, and this was exacerbated by pentoxifylline, the increased penetration rate of hamster oocytes cannot be explained by improved sperm motility. The number of spermatozoa stimulated to acrosome react by 2 μmol A23187 per litre was increased 3‐fold by 3.6 mmol pentoxifylline per litre and 4‐fold by 5 mmol caffeine per litre. These data suggest that CAMP may act synergistically with Ca2+ to stimulate the acrosome reaction. Pentoxifjrlline may improve the fertility of poor‐quality human spermatozoa by enhancing their ability to respond to the Ca2+ signal produced by binding to the zona pellucida.
Superoxide anion production by human spermatozoa as a part of the ionophore‐induced acrosome reaction process
Authors: GRIVEAU J. F. , RENARD P. , LANNOU D. LE
Published: May 2008

The involvement of superoxide anion (O2°‐) in human sperm capacitation and/or acrosome reaction was investigated. Addition of superoxide dismutase (SOD) to the medium at the beginning of the capacitation process or 15 min before induction of the acrosome reaction, decreased the level of ionophore‐induced acrosome reaction. Hyperactivation was unaffected by the presence of SOD during the capacitation process. Addition of calcium ionophore to the sperm suspension increased production of O2°‐ by the spermatozoa by four to five‐fold and induced the acrosome reaction. In the presence of SOD, superoxide anion could not be detected in the medium and the rate of induced‐acrosome reaction was decreased gready. The presence of an inhibitor of protein kinase C inhibited the production of O2°‐ in the medium and reduced the induced‐acrosome reaction. The production of O2°‐ and the acrosome reaction were also increased by exposure of spermatozoa to 12‐myristate 13‐acetate phorbol ester, a specific activator of protein kinase C. While the level of spontaneous acrosome reaction was not increased by the direct addition of O2°‐ to the medium, its presence induced the release of unesterified fatty acids from membrane phospholipids. These findings suggest that the production of O2°‐ by spermatozoa could be involved in the ionophore‐induced acrosome reaction, possibly through the de‐esterification of membrane phospholipids. However, this production of superoxide anion is not sufficient on its own to induce the acrosome reaction.
Quantification of androgen receptor and follicle‐stimulating hormone receptor mRNA levels in human and monkey testes by a ribonuclease‐protection assay
Authors: DANKBAR B. , SOHN M. , NIESCHLAG E. , GROMOLL J.
Published: May 2008

A sensitive, solution‐hybridization ribonuclease‐protection assay (RPA) was established to quantify the expression of mRNA for the androgen receptor (AR) and follicle‐stimulating hormone receptor (FSHR) in total RNA samples isolated from tissues of the cynomolgous monkey, human testes obtained from elderly patients undergoing orchidectomy because of prostatic carcinoma or from transsexual men undergoing gender reassignment as well as human cell lines DU 145, REP and RVP. Sensitivity experiments revealed that, in the human and monkey, 1–2 μg of total RNA were sufficient to achieve quantifiable signals of the different receptor mRNA species. Quantification of AR and FSHR mRNA levels showed a 1.7‐fold higher expression of AR mRNA and a 2.4‐fold higher expression of FSHR mRNA in the monkey testes compared to human testes from patients with prostatic carcinoma. Normal spermatogenesis in both human and monkey testes indicated no relationship between spermatogenic status and receptor expression. The significantly lower expression of AR and FSHR mRNA in humans than in monkeys might therefore be either age‐ or species‐related. Quantification of mRNA for AR and FSHR in the testis of the transsexual patients undergoing oestrogen and antiandrogen treatment displayed a drastic increase (4.5‐fold) in mRNA for the AR, whereas mRNA for the FSHR was barely detectable. Due to its high sensitivity, reproducibility and its ability to quantify mRNA transcripts, the RPA is a useful tool for investigating expression of low abundant receptor genes and their regulation when only very small amounts of tissue are available. Furthermore, it is suitable for use in clinical and experimental studies in which accurate quantification of transcripts is necessary.
Tobacco smoke disrupts testicular microcirculation in the rat
Authors: COLLIN O. , KILTER S. , BERGH A.
Published: May 2008

Anaesthetized adult rats were exposed repeatedly to cigarette smoke for 2 s interspersed with exposure to fresh air for either 10, 15 or 30 s using a smoking apparatus. The acute effects of this treatment on testicular microcirculation were studied using laser Doppler flowmetry. Peripheral tissue O2 saturation was measured continuously in the foot during the experiment. Exposure to cigarette smoke caused a moderate acute increase in testicular blood flow and it inhibited vasomotion. Blood flow and the pattern of vasomotion were normalized after termination of smoke exposure. The time between the last smoke exposure and flow normalization was dependent on the smoke/air dose. Peripheral tissue O2 saturation did not change during the experiments. The effects on testicular microcirculation of local injection of an aqueous extract of cigarette smoke and passive smoking were also studied. Local injection of smoke extract increased blood flow and inhibited vasomotion. Passive smoking caused a moderate decrease in the frequency of vasomotion and amplitude. The observation that cigarette smoke influences testicular microcirculation acutely is discussed in relation to possible adverse effects on spermatogenesis.