bluuebunny/crossref_metadata_2025_split huggingface dataset:
Relative roles of testosterone and the germ cell complement in determining stageldependent changes in protein secretion by isolated rat seminiferous tubules
Authors: Sharpe Richard M. , Millar Mike , Mckinnell Chris 
Published: May 2008

SummaryThis study has compared the effect of withdrawal of testosterone (± replacement) with that of selective depletion of pachytene spermatocytes (PS) or round/elongating spermatids (RS), or both PS and RS, on the level of overall protein secretion by seminiferous tubules (ST) isolated at particular stage‐groups of the spermatogenic cycle. Testosterone withdrawal was induced by destroying the Leydig cells with a single injection of ethane‐dimethane sulphonate (EDS), with or without concomitant replacement of testosterone by injection; ST at stages II‐V, VI‐VIII or IX‐XII were then isolated from control and treated rats at 4 days after treatment. Methoxyaceticacid (MAA) was administered, in either one or two doses, to selectively destroy 80–100% of pachytene and later spermatocytes; ST at stages I‐V, VI‐VIII or IX‐XIV were then isolated at specific times after treatment such that ST were depleted selectively of either PS, RS or PS + RS. Isolated ST (5 cm) were then cultured for 22 h at 34°C with 35S‐methionine and its incorporation into secreted proteins then quantified.Based on the incorporation of 35S‐methionine, ST at stages VI‐VIII showed a significantly higher level of protein secretion than did ST at earlier or later stages. This difference was abolished following testosterone withdrawal but was maintained by testosterone replacement. The normal increase in protein secretion by ST at stages VI‐VIII was also prevented if either PS or RS were depleted, whereas depletion of either PS or RS alone had no significant effect on protein secretion by ST at stages I‐V, and only the depletion of RS significantly reduced protein secretion by ST at stages IX‐XIV. Depletion of both PS + RS reduced protein secretion significantly by ST at all stages. In contrast to the data for total protein secretion, the levels of sulphated glycoprotein (SGP)‐1 and SGP‐2 secreted by ST at stages VI‐VIII showed that these two Sertoli cell proteins were unaffected by germ cell depletion except after co‐depletion of PS + RS when secretion of SGP‐1 was halved. It is concluded that: (i) the increase in overall protein secretion by ST at stages VI‐VIII in the normal spermatogenic cycle is both androgen‐ and germ cell‐dependent: (ii) the modulatory effect of germ cells on ST‐protein secretion varies according to the stage of the spermatogenic cycle and the germ cell type and does not affect all secreted proteins: (iii) the changing germ cell complement is probably the trigger for the stage‐dependent changes in overall protein secretion by ST.
Predictive value of morphologically normal sperm concentration in the medium for in‐vitro fertilization
Authors: ENGINSU M. E. , DUMOULIN J. C. M. , PIETERS M. H. E. C. , EVERS J. L. H. , GERAEDTS J. P. M. 
Published: May 2008

SummaryMorphological evaluation of spermatozoa using strict criteria (MEUSC) and conventional sperm parameters were studied with respect to in‐vitro fertilization and pregnancy outcome before and after a swim‐up selection procedure. Recovered oocytes were inseminated with 50 000 progressively motile spermatozoa, and this study assess the influence of the total number of spermatozoa and of the percentage with strictly normal morphology in the insemination sample on the outcome of IVF. The results showed that the percentages of spermatozoa with normal morphology using strict criteria, both in native and in post‐swim‐up samples, were the best predictors of IVF outcome. Their respective cut‐off points were 5% and 8%. The number of morphologically normal spermatozoa inseminated also showed a good correlation with fertilization. However, it was not possible to find a proper cut‐off point for this parameter. The patients were categorized on the basis of their native and post‐swim‐up scores. Category 1, in which both parameters were below their respective cut‐off points, showed a 7% fertilization rate and a 0% pregnancy rate. Category 3, in which both parameters were above their cut‐off points, showed a 70% fertilization rate and a 23% pregnancy rate. This suggests that sperm morphology can be used as a criterion for patient selection for IVF as an aid to identification of possibly subfertile males.
Paternity in a patient with testicular seminoma and contralateral testicular intraepithelial neoplasia
Authors: DIECKMANN K.‐P. , LOY V. 
Published: May 2008

SummaryA 32‐year‐old patient with unilateral βhCG‐positive seminoma and contra‐lateral testicular intraepithelial neoplasia (TIN; so‐called carcinoma‐in‐situ) with no metastases (clinical stage I) received one course of adjuvant carboplatin therapy. He refused further treatment of TIN in his remaining testis. His wife became pregnant by him 4 months later and delivered a healthy child at term. This case shows that patients with TIN in their remaining solitary testis are not necessarily infertile, and testes afflicted with TIN must also contain tubules that retain normal spermatogenic potential. Surveillance may be an treatment option for patients with TIN in their remaining testis in cases where there is a strong desire for paternity.
Seminal leucocyte subpopulations and sperm function in fertile and infertile Chinese men
Authors: KUNG A. W. C. , HO P. C. , WANG C. 
Published: May 2008

SummaryThe level of seminal leucocytes and the prevalence of leucocytospermia was determined in a group of fertile and infertile southern Chinese men in Hong Kong. Sixteen normal fertile semen donors and 49 men with male factor infertility were studied prospectively. None had antisperm antibodies and past or present evidence of genital tract infection. Seminal leucocytes and their subsets were analysed using monoclonal antibodies and an immunocytochemical alkaline phosphatase‐anti‐alkaline phosphatase conjugate technique. Seminal leucocytes were detectable in 94% and 86% of the fertile and infertile men respectively, with the predominant subset being granulocytes. Leucocytospermia (> 1 × 106 leucocytes/ml) was found in only one of the 49 (2%) infertile men without clinical evidence of genito‐urinary infection. Inverse correlations were observed between (1) the percentage of spermatozoa with normal morphology and the number of T‐helper/inducer cells, (2) the linearity of sperm movement and the number of T‐lymphocytes. In conclusion, the level of seminal leucocytes and the prevalence of leucocytospermia is low in infertile Chinese subjects. The effect of seminal leucocytes on sperm function in these subjects needs further evaluation.
Investigation in real time of the effect of gravitation on human spermatozoa and their tendency to swim‐up and swim‐down
Authors: Makler A. , Stoller J. , Blumenfeld Z. , Feigin P. D. , Brandes J. M. 
Published: May 2008

SummaryTo investigate in real time if and how natural gravity affects rates of swim‐up and swim‐down of human spermatozoa, samples of motile or immobilized spermatozoa in a sealed mini‐chamber were placed vertically on a 90° tilted microscope. The mode of their sedimentation, as well as the difference in the rate of their swimming up and down, were observed directly over 30 min and analysed from photomicrographs. Under the influence of natural gravity force, most immobilized spermatozoa turned their heads down in about 5 min and then sank slowly at an average speed of 0.2 μ/s. The number of motile spermatozoa that swam down was 5–6 times more than those swimming up. It can be implied that in spite of the mild force exerted by 1g on suspended spermatozoa in comparison to the high g force obtained by centrifugation, the overall effect of gravity on the rate of swimming up or down becomes dominant. Gravity causes the sperm heads to turn downward after which the oriented spermatozoa continue to move down by their own tail movements, causing accumulation of motile spermatozoa at the bottom. This may explain why in some recent studies swim‐down was superior to the swim‐up procedure during sperm separation by self‐migration.
Inverse relationship between the induction of human sperm capacitation and spontaneous acrosome reaction by various biological fluids and the superoxide scavenging capacity of these fluids
Authors: Lamirande E. de , Eiley D. , Gagnon C. 
Published: May 2008

SummaryCapacitation of spermatozoa is essential for fertilization, and can be induced by various agents or biological fluids. Previous reports have shown that foetal cord serum (FCS) and the superoxide anion trigger human sperm hyperactivation and capacitation, and that superoxide dismutase (SOD) prevents these processes. We investigated: (1) the capacity of seminal plasma (SP) and follicular fluid (FF)(whole, or fractionated into high and low molecular weight components), in the presence or absence of SOD, to induce the spontaneous acrosome reaction(no stimulant needed, AR) and capacitation (as measured by the lysophosphatidyl‐choline‐induced AR, LPC‐AR); (2) a possible relationship between the levels of AR and capacitation obtained with these biological fluids and the superoxide scavenging capacity of the same fluids. The highest levels of LPC‐AR were obtained with FF ultrafiltrate (48 ± 6%), followed by SPultrafiltrate (31.9 ± 0.8%), FF (30 ± 5%), dialysed FF (27 ± 4%), and finally, by FCSultrafiltrate (23 ± 1%.), SP (21 ± 1%) and dialysed SP (18.9 ± 0.8%).A similar order of potency for the fluids existed when sperm AR was studied, the levels of AR observed ranging from 26 ± 2% to 5.3 ± 0.8% after incubation with FF ultrafiltrate and SP respectively. None of these treatments had detrimental effects on sperm motility. In the presence of SOD, there was always an important reduction (52–86%) of the AR and LPC‐AR observed. A highly significant inverse linear relationship was observed between the SOD‐like activity of the fluids tested and the AR (r= 0.86, p < 0 0.001) and LPC‐AR (r= 0.91, p < 0.001)observed in the presence of these fluids. The results suggest that biological fluids contain inducers of the AR and capacitation, and that these probably act by a common mechanism, possibly by direct or indirect induction of an NADPH oxidase in the sperm membrane. The data suggest strongly that the SOD‐like activity of a specific fluid is probablyone of the most important factors that will determine its capacity to induce the AR and capacitation.
Phenotypic characterization of lymphocytic cell infiltrates into the testes of rats undergoing autoimmune orchitis
Authors: Lustig L. , Lourtau L. , Perez R. , Doncel G. F. 
Published: May 2008

SummaryExperimental autoimmune orchitis (EAO) was induced in adult Wistar rats by active immunization with a testicular homogenate (TH) and adjuvants. Fifty per cent of the immunized rats developed EAO. Testicular damage became evident at 50 days after the primary immunization and increased in severity at 80 days. Phenotypic characterization of T‐cell subsets (CD4+ and CD8+) and Ia+ cells was performed on cryostat sections of testis obtained from normal rats, rats immunized with adjuvant (control group) and rats immunized with TH and adjuvants (experimental group) at 50 and 80 days. Labelled cells were only detected in the interstitial area; no labelled cells were observed inside the seminiferous tubules with any of the monoclonal antibodies used (W3/25, OX‐8, OX‐6). A significant increase in the numbers of CD4+ and CD8+, as well as of Ia+ cells, were observed in the testis of rats with severe EAO at 80 days after the first immunization. Rats of the same experimental group without testicular damage showed no major differences compared to rats from the control group, with the exception of a lower number of CD8+ cells. Variations in the lymphocyte subsets in lymph nodes draining the site of immunization showed the opposite pattern to that observed in the testis. In conclusion, these data suggest the traffic of specifically sensitized lymphocytes from lymph nodes to the testis and an active role of CD4+, CD8+ and Ia+ cells in the pathogenesis of EAO in the rat.
Sperm function in Indonesian men treated with testosterone enanthate*
Authors: ARSYAD K. M. 
Published: May 2008

SummaryThis study was undertaken to assess the fertilizing capacity (sperm function) of residual spermatozoa produced by groups of men (n= 7) rendered oligozoospermic by treatment with weekly intramuscular injection of 50 or 100 mg testosterone enanthate (TE). The treatment was continued for 6 months and the effects compared with a control group (n= 6) in which men received weekly intramuscular injections of sesame oil (1 ml). Sperm function was assessed by tests of membrane integrity (supravital stain and hypoosmotic swelling [HOS] test) and sperm penetration (artificial cervical capillary tube test). The quality of sperm movement was also assessed. These parameters of sperm function were all reduced consistently in a dose‐dependent manner with maximal suppression occurring between weeks 15 and 25 of treatment, and full recovery to baseline levels by 12 months after cessation of treatment. The decrease in sperm function was correlated with the degree of suppression of sperm output with the proportion of morphologically normal spermatozoa and the degree of suppression of gonadotrophin levels. The weekly dose of 50 mg TE induced severe oligozoospermia (concentration < 5 million/ml) in four of seven men, whereas 100 mg TE induced azoospermia in all seven men by week 20. It is concluded that, unlike published observations on men of European background, the administration of TE at 100 mg per week may achieve adequate contraceptive efficacy in Indonesian men.
Effects of retinol on glycoprotein synthesis by Sertoli cells in culture: dolichyl phosphomannose synthase activation
Authors: GUMA FATIMA C. R. , BERNAFCD ELENA AIDA 
Published: May 2008

SummarySertoli cells were isolated from Wistar rats aged 19 days and cultured for 48 h. The addition of retinol (10 μM) to the culture medium significantly stimulated the incorporation of [2–3H]mannose into lipid‐linked oligosaccharide and into cellular and secreted glycoproteins. Incorporation of [U‐ 14C] leucine into proteins and of [5, 6–3 H] uridine into RNA was unaffected by retinol treatment. Incubation of microsomal fractions of retinol‐treated cells showed an increase in mannose incorporation into dolichyl phosphomannose, into dolichyl pyrophosphoryl oligosaccharide and into proteins. Chromatographic analysis of the fraction soluble in chloroform/methanol (2: 1 v/v) did not show the presence of retinyl phosphomannose either in control or in retinol‐treated cells. When the formation of dolichyl phosphomannose was studied in microsomes isolated from control cells and from cells treated with 10 μM retinol for 48 h in the presence of exogenous dolichyl phosphate, the results showed that the retinol effect was due to stimulation of dolichyl phosphomannose synthase.
Outer dense fibres of human spermatozoa: partial characterization and possible physiological functions
Authors: HENKEL R. , STALF T. , MERTENS N. , MISKA W. , SCHILL W.‐B. 
Published: May 2008

SummaryThe outer dense fibres are accessory fibres in the spermatozoon. They represent up to 30% of the protein portion in human spermatozoa and are involved in sperm progressive motility. If outer dense fibres are missing or developed poorly, spermatozoa are only locally motile. For isolation of the outer dense fibres, human spermatozoa were sonicated at 25 kHz and the flagella were separated by density gradient centrifugation in Percoll. Thereafter, membranes and fibrous sheath were dissolved under reducing conditions in the cationic detergent cetyltrimethylammonium bromide for 30, 60 and 90 min, respectively. The isolation steps were monitored by phase contrast microscopy and electron microscopy. After SDS‐polyacrylamide gel electrophoresis and silver staining of isolated outer dense fibres, two protein bands at 55 and 67 kDa could be detected. By means of rhodamine B staining, no phosphorus could be detected in the outer dense fibre proteins.
Flow cytometric evaluation of the acrosome reaction of human spermatozoa: a new method using a photoactivated supravitaI stain
Authors: HENLEY N. , BARON C. , ROBERTS K. D. 
Published: May 2008

SummaryA flow cytometric assay using a double‐stain method for the measurement of the acrosome reaction of human spermatozoa is described. The use of a stable photoactivated stain, ethidium monoazide, allowed evaluation of the viability of spermatozoa. This stain was more stable in fixed samples than propidium iodide, which is not bound covalently to DNA and is therefore removed readily during the washing procedure. The permeabilized acrosome was labelled with Pisum sativum agglutinin conjugated with fluoroisothiocyanate. Since this lectin binds to the acrosome and acrosomal contents, a decrease in the fluorescence intensity allows the cytometric evaluation of the acrosome reaction.Microscopic analysis and flow cytometric analysis were well correlated and cell sorting was performed to ensure the homogeneity of each different subpopulation encountered.
Effects of male accessory sex glands on fertilization, polyspermy and morphometry of early embryos in the golden hamster
Authors: CHOW P. H. , DOCKERY P. , CHEUNG M. l. L. , CHEUNG W. S. O. 
Published: May 2008

SummaryPreimplantation embryos sired by hamsters without accessory sex glands (ASG) were found to have a higher mortality rate and a slower cleavage rate than those sired by sham‐operated males at 72 h post coitum (P.c.). A time‐course study of fertilization in vivo was conducted to determine whether this effect was due to delayed fertilization. Ultrastructural morphometry of 48 h embryos was also undertaken to establish the earliest manifestation of developmental anomalies. Compared to sham‐operated controls (SH), ablation of all the ASG (TX), or just the ventral prostate (VPX) or ampullary gland (AGX) had no effect on the timing of sperm penetration, extrusion of the second polar body and pronuclear formation. Females mated with AGX males tended to have more polyspermic embryos (9.7%; p < 0.05). The volumes, volume fractions (Vv) of the blastomere nuclei, mitochondria and yolk material of the four‐cell embryos sired by these same groups of males were assessed using point counting techniques. No difference in the Vv of yolk and mitochondria could be observed between groups. However, the SH group did have a significantly larger proportion of the cell occupied by the nucleus (p < 0.05), and the TX group had a higher proportion of the nucleus occupied by nucleoli when compared with the SH group (p <0.01). Smaller nuclei and larger nucleoli in the TX group was interpreted as an early manifestation of a slower division rate of the blastomeres.
Clinical study of azoospermia
Authors: MATSUMIYA K. , NAMIKI M. , TAKAHARA S. , KONDOH N. , TAKADA S. , KIYOHARA H. , OKUYAMA A. 
Published: May 2008

SummaryThis study evaluated how many patients with azoospermia might have fertility potential using assisted conception techniques. A total of 102 male patients with aioospermia were included in the study. Thirteen patients had sex chromosomal abnormalities. Testicular biopsy performed in the other 89 patients showed incomplete spermatogenesis in 47 of them whereas 42 had complete spermatogenesis. In the latter 42 patients, distal vasography demonstrated bilateral obstruction of the excurrent ducts in 14 patients whereas no distal obstruction of the ducts was found in 28. The 89 patients were divided into three groups according to the findings of testicular biopsy and distal vasography. In the 14 patients with both complete spermatogenesis and distal obstruction of the excurrent ducts, surgical procedures are applicable. The pathogenesis of the 28 patients with complete spermatogenesis but without distal obstruction of the ducts should be clarified for further treatment.
Post‐thaw sperm motility, cAMP concentration and membrane lipid per oxidation after stimulation with pentoxifylline and platelet‐activating factor*
Authors: WANG R. , BELL M. , HELLSTROM W. J. G. , SIKKA S. C. 
Published: May 2008

SummaryEarlier studies have demonstrated that pentoxifilline (PTX) and platelet‐activating factor (PAF) can significantly improve the motion parameters of post‐thaw human spermatozoa. This study has investigated the effects of PAF, PTX and their combination on cyclic adenosine monophosphate (CAMP) concentrations and membrane lipid peroxidation (LPO) in post‐thaw human spermatozoa. Washed spermatozoa from normal volunteers (n=10) were cryopreserved in Test‐yolk buffer using a standard protocol. After 2 weeks the sperm samples were thawed, washed and incubated with either 1 p˜ PAF, 3 m˜ PTX or 0.5 p˜ PAF plus 1.5 m˜ PTX. Video sequences were recorded at 0, 30, 60, and 120 min for analysis of sperm motion parameters using the Cell Track Sperm Analysis System. Concentrations of cAMP were assessed by radioim‐munoassay, and LPO levels were measured by malondialdehyde‐thiobarbituric acid reactivity. Our studies indicate a time‐course stimulatory effect with overall maximal stimulation observed in samples treated with the combination of PAF and PTX. The maximal stimulation of percentage motility compared to control was observed at 60 min in samples treated with PAR PTX, or PAF plus PTX. PAF plus PTX stimulated straight‐line velocity (VSL), curvilinear velocity (VCL) and lateral head displacement (ALH) after 30 min incubation. The primary effect of PAF was observed on VSL, while the main effect of PTX was on VCL. CAMP concentrations were 3‐fold higher than controls in samples treated with PTX or PAF plus PTX. CAMP concentrations in PAF‐treated samples did not differ significantly from controls. No significant differences were observed between any groups for LPO. This study demonstrates that both PAF and PTX can improve the motion parameters of post‐thaw human spermatozoa with the maximal effect observed by their combination. The stimulatory effect of PTX in these post‐thaw samples is related to increased CAMP levels, while another mechanism is likely to be operational for PAR Protection from lipid peroxidative membrane damage is not suggested to be the mechanism of action for PAF or PTX under these conditions.
Sperm motility inhibitor from human seminal plasma: presence of a precursor molecule in seminal vesicle fluid and its molecular processing after ejaculation
Authors: ROBERT MARTIN , GAGNON CLAUDE 
Published: May 2008

SummaryHuman seminal plasma contains a protein factor that has the capacity to inhibit the movement of demembranated and intact spermatozoa. This factor ‘seminal plasma motility inhibitor’ (SPMI) has been shown to originate exclusively from the seminal vesicles. The present results demonstrate that the biological activity of SPMI in semen decreases rapidly from 1000 U/d, immediately after ejaculation, to 220 U/ml 2 h later. Immunoblots of seminal plasma proteins probed with an antibody against human SPMI, revealed the rapid processing of a predominant 52 kDa SPMI antigen, present in the seminal vesicle secretions. This precursor was degraded initially into intermediate molecular mass fragments of 25–40 kDa, and subsequently into smaller fragments of 17–21 kDa. When seminal vesicle fluid was mixed with prostatic secretions (3: 1 v/v), proteases present in prostatic secretions were shown to be responsible for processing of the SPMI precursor. Addition of protease inhibitors such as phenylmethylsulphonyl fluoride (PMSF, 5 mM), benzamidine (100 mM) or ethylenediaminetetraacetic acid (EDTA, 5 mM) to the mixture of seminal vesicle and prostate secretions partially prevented the loss of activity of SPMI by 54%, 27% and 9%, respectively. However, the simultaneous addition of PMSF and benzamidine conferred almost total stability to the SPMI precursor activity. These results demonstrate that SPMI exists as a predominant 52 kDa precursor form in the seminal vesicles and is processed rapidly after ejaculation into less active, lower molecular mass forms by one or more serine proteases and/or metallo‐proteases of prostatic origin which are present in liquefied semen.