bluuebunny/crossref_metadata_2025_split huggingface dataset:
Induction of spermatogenesis with gonadotrophins in Chinese men with hypogonadotrophic hypogonadism
Authors: KUNG A. W. C. , ZHONG Y Y. , LAM K. S. L. , WANG C. 
Published: May 2008

SummaryThe effects of gonadotrophin administration to 17 Chinese patients with hypogonadotrophic hypogonadsm (HH) on testicular volume and induction of spermatogenesis were studied. Ten subjects had isolated HH and seven had hypopituitarism. Twelve of the subjects had prepubertal onset of HH and five of them had been treated previously with hCG for induction of puberty. None had a history of cryptorchidism. During hCG treatment for induction of spermatogenesis, all subjects had an increase in serum levels of testosterone into the normal adult male range and their testes increased in size from 3 (1–20) ml to 11.6 (5–20) ml [median(range), p<0.02]. Six subjects required treatment with hCG alone. However, the remaining 11 subjects, after at least 6 months treatment with hCG, required the addition of human menopausal gonadotrophin (hMG) to induce spermatogenesis. Two subjects remained azoospermic. One had a history of mumps orchitis and the other had isolated elevation of blood FSH levels, suggestive of primary testicular failure in addition to HH. Excluding one subject with fertile eunuch syndrome, the mean duration for first appearance of spermatozoa was 13 (4–52) months. Twelve subjects became fertile and pregnancy was achieved in their partners after 20 (4–78) months. The weekly doses for hCG and hMG were 4000 (3000–10000) IU and 225 (225–450) IU, respectively. Patients who responded to hCG alone had a significantly larger pretreatment testicular volume, suggesting that they had only partial gonadotrophin deficiency. Prepubertal onset of hypogonadism was not a determining factor for requirement of hMG treatment. Pretreatment testicular volume correlated positively with the final testicular volume and negatively with the time to achieve spermatogenesis, but not with the final sperm concentration. Previous use of hCG for induction of puberty with resultant testicular growth, favoured a subsequent positive response to hCG. These subjects also required a lower dose of hCG for normalization of serum testosterone levels. In conclusion, gonadotrophins are very effective in stimulating testicular growth and spermatogenesis in subjects with HH, and the positive response to gonadotrophins can be determined by the pretreatment testicular volume.
Morphometric analysis of human sperm morphology
Authors: PÉREZ‐SÁNCHEZ F. , DE MONSERRAT J. J. , SOLER C. 
Published: May 2008

SummaryFourteen morphological forms of human spermatozoa were analysed morphometrically using semi‐automated image analysis techniques. Five basic (area, perimeter, length, width and mass) and five derived (ratio, length minus width, ellipticity, form and total mass) parameters were considered. Statistical analysis showed differences among all 14 types of human sperm heads. Basic parameters describing the size and shape were enough to distinguish most of the categories, whereas derived parameters as well as parameters dependent on stain intensity, were demonstrated to be useful for the discrimination of some morphological categories. The fact that statistical analysis showed differences among all 14 sperm types provides evidence for the reliability of our morphological classification. These results show that morphometry can be used for the fine study of sperm morphology and may serve as a database for future work dealing with sperm classification. As this is a pilot study to assess methodology, further studies will be required to validate the method in terms of its application and usefulness in assessing the fertilizing potential of human spermatozoa.
The routine assessment of sperm motility at room temperature and 37°C
Authors: BIRKS A. G. , IZZARD H. , MORROLL D. R. , PRIOR J. R. , TROUP S. A. , LIEBERMAN B. A. , MATSON I. L. 
Published: May 2008

SummaryThe World Health Organization (WHO) laboratory manual (1992) states that assessment of sperm motility can be performed at either 37OC or room temperature (20–24OC). The motility of spermatozoa in 44 semen samples (22 fresh samples and 22 frozen‐thawed samples) was assessed at both of these temperatures and a significant difference in the motility profiles was noted, specifically an increase at 37OC in the percentage (expressed here as median and ranges) of spermatozoa with excellent progressive motility and an overall increase in the percentage with total progressive motility. With fresh samples the excellent progressive motility increased from 41 (19–53) to 54 (30–66) and the overall motility from 58.5 (39–74) to 65.0 (40–79). With the frozen—thawed samples the excellent motility increased from 14 (1–33) to 25 (6–45) and the overall motility from 30.5 (14–51) to 33.0 (16–52). As the WHO laboratory manual was published. ‘In response to a growing need for the standardisation of procedures for the examination of human spermatozoa’ it is proposed that only one temperature for routine analysis should be used, namely 37OC, which may have more physiological relevance and eliminate effects of fluctuations in ambient laboratory temperature.
Spermatogenic arrest and ‘Sertoli cell‐only’ syndrome—common alcohol‐induced disorders of the human testis
Authors: PAJARINEN J. T. , KARHUNEN P. J. 
Published: May 2008

SummaryThe effect of alcohol on spermatogenesis and morphometric analysis of the human testis was investigated in a prospective autopsy study, with detailed alcohol‐use reports from relatives or friends of the deceased. The autopsy series comprised non‐alcoholic controls (daily intake <10 g; n=32) and heavy‐drinkers (n=44) with an intake of >80 g per day. Of the controls, 26 (81.3%) men showed normal spermatogenesis, and six (18.7%) partial spermatogenic arrest. Of the heavy drinkers, only 16 (36.4%) had normal spermatogenesis (p<0.00l), while 23 (52.3%) showed partial or complete spermatogenic arrest (p<0.00l) and five had Sertoli cell‐only (SCO) syndrome (pC0.05). The mean testicular weight of heavy‐drinkers was slightly (p<0.05) lower than in the controls. Compared to men with normal spermatogenesis, testicular weight was slightly lower both in controls and heavy‐drinkers with spermatogenic arrest, and was significantly (p<0.01) lower in heavy‐drinking men with SCO syndrome. Spermatogenic arrest was not correlated with fatty liver or cirrhosis of the liver, whereas four of the five men with SCO syndrome exhibited a fatty liver. Thus, our results suggest that the most common alcohol‐related pathological change in the testes is probably reversible arrest of spermatogenesis. Furthermore, we suggest that many heavy social drinkers may suffer from self‐inacted infertility, as one tenth of heavy‐drinkers in our study had SCO syndrome.
Regional variation of specific gene expression in the dog epididymis as revealed by in‐situ transcript hybridization
Authors: PERA I. , IVELL R. , KIRCHHOFF C. 
Published: May 2008

SummaryIn‐situ transcript hybridization was used to characterize the regional distribution of three marker gene transcripts which are expressed abundantly in the canine epididymis. The gene products CE1, CE4 and CE5, which are the canine equivalents of the human homologues HE1, HE4 and HE5, are shown to be expressed in a tissue‐specific and regionally characteristic pattern in the epicfidymal epithelium. CE1 mRNA was expressed very weakly in the efferent ducts but was expressed at a high level throughout the caput and corpus regions of the epididymis, decreasing somewhat in the distal cauda region. CE4 mRNA was not detectable in the efferent ducts and was only expressed moderately in the remainder of the epididymis, with greatest levels in the caput and proximal cauda regions, and decreasing in the distal cauda. CE5 mRNA showed the most marked regional variation in levels with little or no mRNA detectable in the caput and proximal corpus regions, but increasing dramatically in the distal corpus and cauda. In the transition region of the central corpus, the CE5 mRNA appeared to be expressed intermittently, giving a mottled signal appearance over the epididymal epithelium. The patterns of mRNA distribution for the three marker genes in the dog epididymis were, therefore, essentially similar to those for the equivalent human homologues, providing further support for the suitability of the dog epididymis as a model for the human.
Proteins in fluid from the proximal vas deferens of normal fertile and vasectomized men
Authors: FENG H.‐M. , WEN R.‐Q. , LI S.‐Q. , WANG C. , WANG Q.‐H. , JIANG Y.‐J. , LI Q.‐K. 
Published: May 2008

SummaryFluids were collected from the proximal vas deferens of 18 normal fertile men and 32 vasectomized men during vasectomy or vasovasostomy, respectively, and the protein concentration and pattern of proteins were then analysed by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS‐PAGE). In normal fertile men and vasectomized men there were no significant differences in total protein concentrations between fluids from the left and right vas deferens. The total protein concentration of proximal vas fluid from vasectomized men (82.8 ±7.5 mg/ml; mean ± SEM) was significantly higher than in normal fertile men (36.8 ± 4.5 mg/ml). In vasectomized men there was no significant correlation between the total protein concentration in vas fluid and the duration of vasectomy. The patterns of protein bands in proximal vas fluid differed from those in seminal plasma. There was no relationship between the pattern of protein bands in vas fluid and the duration of vasectomy. Five major groups of proteins from proximal vas fluid were identified and no new major proteins were found in proximal vas fluid after vasectomy. The percentage of 80K and 67K protein bands in vasectomized men was higher than that in normal fertile men. It is suggested that, after vasectomy, the physiological environment of the proximal vas deferens may be changed. One of these changes may be the higher total protein concentration (especially the 80K and 67K proteins) in vas fluid.
In vitro fertilization and the effect of progesterone and 17α‐hydroxyprogesterone on acrosome reaction of mouse epididymal spermatozoa
Authors: KHOLKUTE S. D. , RODRIQUEZ J. , DUKELOW W. R. 
Published: May 2008

SummaryThis study assessed the effect of progesterone and 17α‐hydroxyprogesterone (17α‐OH‐progesterone) at concentrations of 0.01–10 μg/ml, on the acrosome reaction and in vitro fertilizing ability of mouse epididymal spermatozoa. Cumulus masses containing oocytes were cultured in Brinster's medium, to which were added capacitated epididymal spermatozoa which had been incubated in medium with various concentrations of progesterone or 17α‐OH‐progesterone for 90 min. IVF success rate was assessed 20–24 h following insemination. Progesterone was found to increase the fertilization rate at the 1 μg/ml and 10 μg/ml concentrations while lower concentrations had no effect. However, 17α‐OH‐progesterone failed to show any effect on fertilizing ability. Incubation of epididymal spermatozoa in medium containing 1 μg/ml and 10μg/ml progesterone significantly increased the acrosome reaction as monitored by a chlortetracycline fluorescence assay. 17α‐OH‐progesterone, however, failed to show any effect on the acrosome reaction. The results suggest direct effects of progesterone, but not of 17α‐OH‐progesterone, on fertilization and the acrosome reaction of mouse spermatozoa.
Functional and structural characteristics of human epididymal spermatozoa retrieved by transcutaneous aspiration
Authors: FORESTA C. , ZORZI M. , GALEAZZI C. , ROSSATO M. 
Published: May 2008

SummaryThe present paper describes the development of a simple, repeatable technique for transcutaneous aspiration of epididymal spermatozoa in patients with inoperable excretory azoospermia. Some functional and structural parameters of the retrieved spermatozoa were also evaluated. Epididymal spermatozoa from patients and ejaculated spermatozoa from fertile donors showed similar percentages of acrosome‐reacted cells after incubation for 6 or 15 h in capacitating medium. Penetration of zona‐free hamster oocytes, expressed as the percentage of penetrated ova, was significally lower (p < 0.01) for spermatozoa from patients than for ejaculated spermatozoa from fertile donors. The percentage of epididymal spermatozoa with stable nuclei in sodium dodecyl sulphate was lower (p < 0.001) than for ejaculated spermatozoa from fertile donors. A positive linear correlation (r= 0.74, p < 0.005) was evident between the oocyte penetration rate and the percentage of spermatozoa with stable nuclei. In conclusion the percutaneous retrieval of epididymal spermatozoa in excretory azoospermia allows collection of motile spermatozoa, which can be evaluated for their structural and functional characteristics before repetition of the technique for retrieval of spermatozoa for assisted fertilization techniques.
Characterization of development‐specific, cell type‐specific mouse testicular antigens using testis‐specific polyclonal antibodies
Authors: TSUCHIDA J. , NISHINA Y. , AKAMATSU T. , NISHIMUNE Y. 
Published: May 2008

SummaryAntisera were raised by immunizing rabbits with mouse testicular germ cells, and absorbed in vivo by injection into castrated male whole mice to obtain a specific antiserum which reacted with mouse germ cells. The expression of mouse testisspecific antigenic macromolecules was then studied immunochemically with the antiserum. Approximately 20 antigenic macromolecules with molecular weights ranging from 26 to 110 kD were detected in the normal adult testis. At least 12 of these were differentiation‐specific antigens appearing during development of germ cells, while others were expressed in testicular germ and/or somatic cells or detected only in mature spermatozoa. This technique for raising testis‐specific antisera could be useful for isolation of cDNA clones encoding their antigens, as well as for investigation of the physiological roles of these molecules in germ cell differentiation at the molecular level.
Influence of batches of bovine oocytes on the outcome of an intact zona pellucida binding assay and in vitro fertilization
Authors: ZHANG B. R. , LARSSON B. , RODRIGUEZ‐MARTINEZ H. 
Published: May 2008

SummaryA series of experiments was performed in vitro to determine the variation between batches of abattoir‐derived bovine oocytes to bind and to be fertilized by frozen–thawed spermatozoa provided by a single ejaculate from a bull with high field fertility after artificial insemination. Fourteen batches of oocytes, collected over a 90‐day period, were divided randomly into two equal groups at each sampling period. One group of oocytes was subjected to in‐vitro maturation and subsequently in‐vitro fertilization (IVF) while the other group was used as intact oocytes in a zona binding assay (ZBA). Fertilized oocytes were evaluated for penetration and pronuclear formation and, in the ZBA, the number of spermatozoa bound to each oocyte was registered. Neither the results of the ZBA nor the IVF results varied significantly (p=0.45) between batches. Furthermore, when a sperm concentration of 0.5 × 106/ml was used, the number of spermatozoa bound to the zona pellucida correlated significantly with the frequency of oocytes penetrated after IVF (r= 0.76). The results suggest that variation between the batches of oocytes has no detrimental significance for the application of intact zona pellucida binding or IVF for assaying frozen–thawed bull semen, provided that a large number of oocytes/batch (> 15) are assayed.
Functional motion changes during sperm transit to the site of fertilization and in‐vitro applications: a review
Authors: MBIZVO M. T. 
Published: May 2008

SummaryContinued research to define the parameters of sperm function should aid the evaluation of various approaches in infertility management as well as the efficacy of contraceptives for men which do not necessarily achieve azoospermia.Many treatment forms have been advocated for male factor infertility but have yielded little effect. These include, for example, gonadotrophins, clomiphene citrate, the weakly androgenic steroid, mesterolone. Often, improvements in oligoasthenozoospermia that are not related to genital infection, do not attain normozoospermic levels.Owing to lack of success with the various treatment modalities, assisted reproductive technology encompassing artificial insemination by husband or donor following in vitro enhancement of sperm function have assumed an important role in male infertility. Agents that have been shown to induce and support sperm capacitation processes such as hyperactivation, could serve an important role. These include human follicular fluid (HFF), maternal serum, fetal cord serum and methyl xanthine derivatives.
Sperm binding to the human zona pellucida after migration through human cervical mucus
Authors: VIGIL P. , RIQUELME R. , MORALES P. 
Published: May 2008

SummaryDuring lactational amenorrhea a special type of cervical mucus, similar to that found during the luteal phase, is produced. This mucus, however, is able to support sperm migration. In the study described, the ability of spermatozoa to bind to the human zona pellucida (hZP) after migration through periovulatory and post‐partum mucus was studied. Mucus was obtained from exclusively breastfeeding women in amenorrhea at 30, 60, 120 and 180 days post‐partum. Periovulatory mucus samples from normally cycling women were used as a control. Flat capillary tubes were filled with BWW culture medium at the top and cervical mucus at the bottom. The tubes were immersed in a semen reservoir and the spermatozoa allowed to migrate through the mucus for 3 h into the culture media. Then the spermatozoa were coincubated with 3–4 hZP for 30 min and the number of bound spermatozoa per zona was counted. Periovulatory cervical mucus had an average Insler score of 14 ± 0.5 as compared to 4.6 ± 0.4 for post‐partum mucus. Spermatozoa recovered from periovulatory mucus were always able to bind to the hZP. Spermatozoa recovered from post‐partum mucus, however, were able to bind to the hZP in only 68 ± 7% of the cases. Moreover, spermatozoa recovered from post‐partum mucus bound to the ZP in lower numbers than did spermatozoa recovered from periovulatory mucus (p<0.03). These results suggest a greater ability of sperm‐hZP binding after migration through periovulatory mucus and they also indicate that sperm binding to the ZP is possible even after sperm migration through a low quality mucus.
Assessment of the vitality and acrosomal status of human spermatozoa using fluorescent probes
Authors: KINGER S. , RAJALAKSHMI M. 
Published: May 2008

SummaryThe acrosomal status and vitality of human spermatozoa are generally assessed simultaneously using the lectins Pisum sativum agglutinin (PSA), Peanut agglutinin (PNA) and Concanavalin A (Con A) in conjunction with either Hoechst 33258 (H258; a fluorescent DNA‐binding supravital stain with limited membrane permeability) or a hypo‐osmotic swelling (HOS) test. In the present study, sperm vitality was assessed using H258 under different staining conditions and compared with that estimated using eosin‐nigrosin (E‐N) under different staining conditions and the HOS test. The sensitivity of PNA‐ and Con A‐labelling methods were also compared by evaluating the acrosomal status of motile human spermatozoa after capacitation (1 and 4 h) in vitro followed by exposure to dimethylsulphoxide (DMSO) and a calcium ionophore. The E‐N method employing eosin‐staining for 15 sec, rather than for 30 sec, provided a more reliable estimate of sperm vitality. H258, as used in the H258/Con A‐labelling method (with and without ethanol fixation) rather than under the staining conditions equivalent to H258/PNA‐labelling, was as good for vitality assessment as the E‐N method employing eosin‐staining for 15 sec. However, the HOS test overestimated the proportion of dead spermatozoa compared to those obtained using different H258 and E‐N methods. Further, the Con A‐labelling method consistently scored a significantly lower percentage of spermatozoa undergoing the acrosome reaction compared to those estimated by the PNA‐labelling method. It is concluded that the different sensitivity of these methods can be attributed to the different binding specificity of the lectins. Since PNA and Con A specifically label the outer and inner acrosomal membrane, respectively, spermatozoa in the early stages of the acrosome reaction, undergoing modification of the outer acrosomal membrane but retaining their acrosomal contents, would be scored as acrosome‐reacted by the PNA‐labelling method but as acrosome‐intact by the Con A‐labelling method. The lower sensitivity of the Con A‐labelling method could also be due to the use of aldehyde‐fixed spermatozoa for Con A‐labelling. Also, the PNA‐labelling requires less time to complete than does the Con A‐labelling.
A simple method to detect sperm chromatin abnormalities: cytochemical mechanism and possible value in predicting semen quality in assisted reproductive procedures
Authors: ANDREETTA A. M. , STOCKERT J. C. , BARRERA C. 
Published: May 2008

SummaryProtamine crosslinking by disulphide (‐SS‐) bonds is the main factor responsible for the stability of chromatin structure in mammalian spermatozoa. Sperm chromatin containing arginine/cysteine‐rich protamines shows a deeply modified cytochemical reactivity (e.g. basophilia) when compared with somatic chromatin. After methanol or ethanol‐acetic acid fixation and toluidine blue (TB) staining, most sperm heads in semen smears from human fertile donors exhibited a pale blue (orthochromatic) colour, while a few sperm heads exhibited violet‐blue or violet (metachromatic) staining. Smears from infertile patients showed an increased amount of metachromatic sperm nuclei. After reduction of ‐SS‐ bonds by dithiothreitol, sperm heads from all smears were metachromatic, suggesting that DNA phosphates then become available for TB stacking. Micro‐ and macro‐spectrophotometric studies confirmed the microscopic colour reaction of sperm nuclei. The ortho‐/metachromatic ratio seems a useful parameter for evaluation of altered chromatin structure in spermatozoa cells. Taking into account the current interest in complementary staining tests for evaluation of semen quality, the metachromatic TB reaction represents a simple cytochemical approach for detecting sperm chromatin abnormalities based on differences in ‐SS‐ crosslinking.
Intravasal injection of formed‐in‐place medical grade silicone rubber for vas occlusion
Authors: SOEBADI D. M. , GARDJITO W. , MENSINK H. J. A. 
Published: May 2008

SummaryThis paper describes two consecutive studies: a volume study and an efficacy study. The volume study determined the appropriate volume of Medical Grade Silicone Rubber (MSR) needed to achieve complete occlusion of the vas deferens. This was done by in‐vitro testing of 130 human vas specimens containing plugs of MSR formed in vivo. The volume of MSR needed to occlude the vas was 0.1531 ± 0.0059 ml injected by five to six turns of the applicator handwheel. There was a correlation between MSR volume, weight, and the number of turns of the applicator handwheel. An influence of body height upon the volume and weight of the MSR was also observed. An oval shaped clamp (15 mm long) was more effective in producing secure vas occlusion than was a round clamp (10 mm long). The mean outer diameter of the vas specimens was 1.80 ± 0.15 mm, and the mean maximum dilated inner diameter was 0.93 ± 0.11 mm.The efficacy study was a clinical trial to compare MSR vas occlusion (using an oval 15 mm clamp and the appropriate volume derived from the volume study, n=58) with no‐scalpel vasectomy as the standard procedure (n=64). The azoospermia rate following MSR vas occlusion was not significantly different from that following no‐scalpel vasectomy, and was achieved in 3–6 months.