bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantitative evaluation of the maintenance and development of spermatocytes and round spermatids in cultured tubule fragments from immature rat testis
Authors: TOEBOSCH A. M. W. , BRUSSÉE R. , VERKERK A. , GROOTEGOED J. A. 
Published: May 2008

SummaryMaintenance and development of spermatocytes and round spermatids was studied in an in‐vitro incubation system. This system consisted of open tubule fragments from 26–day‐old rat testes, obtained after collagenase treatment. The tubule fragments contained Sertoli cells and spermatogenic cells up to and including a small number of early round spermatids. The number of primary spermatocytes and round spermatids in the tubule fragments was estimated using flow‐cytometric analysis, immediately after isolation and after 72 h of incubation. In addition, the activity of LDH‐C4 in the tubule fragments was measured. After 72 h of incubation, the percentage of spermatocytes was reduced by 70–80%, but the percentage of spermatids was doubled. The total LDH‐C4 activity per well was increased 2–3–fold during 72 h of incubation of the fragments. A modest improvement of the culture results was observed when a combination of FSH, insulin, retinol and testosterone was added to the medium. LDH‐C4 activity was investigated to see whether it could be used as a quantitative marker of isolated and cultured spermatocytes and spermatids. It was observed that LDH‐C4 activity per cell was decreased when spermatocytes and spermatids were isolated and/or incubated at 4°C. However, the cellular enzyme activity returned to control values during subsequent incubation of the cells at 32°C, either in the absence or presence of a protein synthesis inhibitor. Cellular LDH‐C4 activity may be influenced not only by temperature, but possibly also by other cell isolation conditions. It is concluded that LDH‐C4 activity may not be a reliable quantitative marker for the presence of spermatocytes and spermatids in culture, but should be used in combination with other analytical methods such as DNA estimation and DNA flow cytometry.
Variation in, and inter‐relationship between, prostaglandin levels and other semen parameters in normal men
Authors: SVANBORG K. , GOTTLIEB C. , BENDVOLD E. , YGDEMAN M. B. 
Published: May 2008

SummaryIn each of 135 ejaculates from 46 healthy men, semen volume, various sperm parameters and the level of prostaglandins (PGs) were determined. Data were analysed to determine the within‐ and between‐subject variability, the effect of abstinence period and the correlation between different parameters. Data which did not reveal a Gaussian distribution were transformed logarithmically. For all variables the between‐subject variance exceeded the variation within subjects (P < 0.001). Variation in time of abstinence between 1 and 3 days influenced semen volume and sperm content as well as the content of 19‐hydroxylated PGs, but did not affect sperm motility or the concentration of PGE or PGF. A significant negative correlation was found between the seminal content of PGE and PGF and the total sperm count as well as the sperm concentration. Sperm motility was related to the balance between the content of 19‐hydroxy PGE and 19‐hydroxy PGF but not to their separate concentrations. The ratio between 19‐hydroxy PGE and 19‐hydroxy PGF did not change when the time of abstinence differed. Synthesis of the two 19‐hydroxylated PGs seemed to be regulated by the same mechanism, but their hydroxylation capacity still varied considerably between individuals.
Purification of an acrosomal antigen recognized by a monoclonal antibody and antifertility effects of isoimmune serum
Authors: LIU M.S. , YANG Y. , PAN J. , LIU H.‐W. , MENGE A. C. , LEE C.‐Y. G. 
Published: May 2008

SummaryA highly conserved acrosomal antigen reactive to a monoclonal antibody (HS‐63), generated against human sperm, was purified to homogeneity with a combination of conventional procedures and immunoaffinity chromatography using a soluble extract of mouse and rabbit testes. The molecular weight of the purified antigen was 42–50 kD when analysed by sodium dodecylsulphate polyacrylamide gel electrophoresis. The high specificity of the purified antigen to monoclonal antibody HS‐63 was shown by indirect immunofluorescent inhibition assay, enzyme‐linked immunosorbent assay, Western blot analysis and radioimmunosorbent assay. The purified antigen was used for isoimmunization of mice and rabbits. Following successive immunizations, antisera of high titres were raised and reacted specifically with antigen on the sperm acrosome and in testes of several mammalian species, but not with somatic tissues. These isoimmune sera exhibited strong inhibition on mouse in‐vitro fertilization and human sperm penetration of zona‐free hamster eggs. The results of this study suggest that the sperm‐specific acrosomal antigen reacting with HS‐63 could be a good candidate for the development of immunocontraceptive vaccines in humans and in other animals.
Presence and distribution of leucocyte subsets in the murine epididymis after vasectomy
Authors: NASHAN D. , COOPER T. G. , KNUTH U. A. , SCHUBEUS P. , SORG C. , NIESCHLAG E. 
Published: May 2008

SummaryMale mice were vasectomized by ‘open‐ended’ or ‘closed’ techniques. After 4 weeks the cell‐mediated immune reactions were compared with those of sham‐operated animals by immunohistochemical localization of leucocytes, using specific monoclonal antibodies. Macrophages and MHC class II antigen‐positive cells were the major cell types to appear in all regions of the epididymis after both types of operation. There was recruitment of T‐helper/inducer leucocytes but not of T‐suppressor‐cytotoxic cells. An increased presentation of macrophage‐migration inhibiting factor antigen appeared in interstitial and peritubular locations. After ‘closed’ and ‘open‐ended’ vasectomy granulomata developed in the epididymis. The sperm‐containing lumen of these granulomata was invaded by macrophages, MHC class II‐positive cells and T‐helper/inducer lymphocytes. This mouse model thus reveals a significant epididymal inflammatory response of the epididymis to vasectomy.
Laminin, type IV collagen, and fibronectin in normal and cryptorchid human testes. An immunohistochemical study
Authors: SANTAMARIA L. , MARTINEZ‐ONSURBE P. , PANIAGUA R. , NISTAL M. 
Published: May 2008

SummaryAn immunohistochemical study of laminin, type IV collagen, and fibronectin was carried out in the testes of normal men and in the cryptorchid and contralateral scrotal testes of cryptorchid men from 2 to 40 years of age. The integrated optical density (IOD) per unit area of the lamina propria was measured in the immuno‐stained sections. Fibronectin was found throughout the thickness of the lamina propria of the seminiferous tubules and in the interstitial connective tissue. No differences between normal and cryptorchid testes were found. Laminin was observed in the innermost part of the lamina propria of the seminiferous tubules and surrounding the endothelium of blood capillaries from infancy. No differences were found between normal and cryptorchid testes in the prepubertal period. In adult cryptorchid testes, laminin formed more numerous and deeper invaginations towards the seminiferous epithelium than in normal adult testes. Type IV collagen appeared throughout the thickness of the lamina propria of normal testes as well as in the wall of interstitial blood vessels. From infancy, the lamina propria of seminiferous tubules, but not blood vessel walls, showed lesser immunostaining for type IV collagen and a lower IOD of this component than did control tests from men of the same age. No differences between unilateral and bilateral cryptorchid‐ism were found. The contralateral scrotal testes of cryptorchid males showed intermediate immunostaining for type IV collagen between that of normal control testes and that of cryptorchid testes. These findings suggest that the lamina propria of seminiferous tubules is lesioned at an early age in both cryptorchid and contralateral scrotal testes of cryptorchid men.
Cloning and analysis of mRNAs expressed specifically in the human epididymis
Authors: KIRCHLOFF C. , OSTERHOFF C. , HABBEN I. , IVELL R. 
Published: May 2008

SummaryDifferential cDNA screening identified four new genes that are expressed specifically in the human epididymis. Their corresponding mRNAs are relatively short, ranging from 0.7 to 1.0 kb, and belong to a class of abundant or moderately abundant transcripts. Tissue‐specific expression was pronounced for all four sequences, which showed no hybridization signals with RNA from eight different human tissues. Cross‐hybridization with epididymal RNAs from some non‐rodent mammalian species was observed with two cDNAs; the remaining two transcripts failed to cross‐hybridize with any of the animal epididymal RNAs tested.
Age‐related differences in testicular microcirculation
Authors: DAMBER J. ‐E. , BERGH A. , WIDMARKt A. 
Published: May 2008

SummaryDifferent aspects of testicular microcirculation were studied in rats aged 20, 24 or 90 days, in order to determine if maturational changes in the testis could influence its vasculature. Using laser Doppler flowmetry, it was found that vasomotion was not present in 20‐day‐old rats, but it could be induced by hCG‐treatment given 32 h prior to flow measurements. In rats aged 24 days vasomotion was present in the testis but it was not influenced by hCG treatment, in contrast to adult rats in which testicular vasomotion disappeared after hCG treatment. In all age‐groups, hCG treatment induced an increase in testicular interstitial fluid (IF) volume, which was most pronounced in young rats. In young rats, as in adults, there was an increase in the volume density of polymorphonuclear (PMN) leucocytes in testicular blood vessels after hCG treatment. However, rats aged 20 or 24 days did not exhibit any significant extravascular migration of leucocytes as occurred in adult rat testis after hCG injection. The deposition of colloidal carbon particles observed in adult rats after hCG‐treatment was also not seen in young rats. It is concluded that the vasculature of the immature rat testis reponds to hCG stimulation with an increase in IF volume. However, when compared to adult rats, different mechanisms may be operating because no evidence for opening of inter‐endothelial cell gaps was observed in the testis of young rats.
Anti‐spermatogenic activity of malvidin chloride in langur monkeys (Presbytis entellus entellus Dufresne)
Authors: BHARGAVA S. K. 
Published: May 2008

SummaryMalvidin chloride (MC) a colouring agent from flowers of Malvaviscus conzattii Greenum was studied for male anti‐fertility effects in adult langur monkeys (Presbytis entellus entellus Dufresne). When fed 50 mg MC kg ‐1 for a period of 60 days, inhibition of spermatogenesis resulted. The weights of testes and epididymides were reduced and there was atrophy of the Leydig cells. In the epididymis, epithelial cell heights were reduced after MC‐treatment. Conspicuous shrinkage of seminiferous tubules and Leydig cell nuclei were evident. Depletion of total proteins, RNA, sialic acid, alkaline/acid phosphatase activity in testes and epididymides with the elevation of testicular levels of cholesterol and glycogen also occurred. Blood/serum analysis and haematology of MC‐treated langur monkeys revealed that all of the clinically important parameters were well within the normal range. The slightly increased bilirubin concentration and alkaline phosphatase activity returned to normal range within 30 days of the last dose of MC. The anti‐spermatogenic activity of MC in langur monkeys is discussed.
The levels and possible involvement of leukotriene B4 and prostaglandin F2α in the control of interstitial fluid volume in the rat testis
Authors: ABAYASEKARA D. R. E. , KURLAK L. O. , JEREMY J. Y. , DANDONA P. , SHARPE R. M. , COOKE B. A. 
Published: May 2008

SummaryThe possible involvement of two arachidonic acid metabolites, prostaglandin F2α (PGF2α) and leukotriene B4 (LTB4), in the stimulatory effect of human chorioni gonadotrophin (hCG) on the volume of interstitial fluid (IF) in the rat testis ha been investigated. Administration of hCG caused a time‐ and dose‐dependen increase in the IF levels of PGF2α while LTB4 showed no clear dose‐dependence but did decrease significantly at 2–8 h after injection of 100 IU hCG. Administra tion of ethane dimethane sulphonate (EDS), which specifically destroys Leydi cells, decreased the volume of IF but the IF levels of LTB4 and PGF2α were unchanged. This indicates that although the absence of Leydig cells results i significant changes in IF volume, LTB4 and PGF2α are probably not involved in these changes. Furthermore, these findings suggest that Leydig cells are not th only contributors to LTB4 and prostaglandins E2 and F2α in testicular IF. In rats injected peripherally with hCG, intratesticular administration of the cyclo oxygenase inhibitor, indomethacin, decreased PGF2α, levels in IF after 2 and 4 ’ but not at later times. However, no inhibitory effect of indomethacin on the hCG stimulated increase in IF volume was detected; indeed, at 2 h after injection o indomethacin + hCG there was a significant increase in IF volume in the indomethacin‐injected testis. It is concluded that, although prostaglandins am LTB4 are secreted into IF in the rat testis, these arachidonic acid metabolites ar apparently not the primary mediators of the increase in IF volume that follows hCC treatment.
Analysis of the relationship between reactive oxygen species production and leucocyte infiltration in fractions of human semen separated on Percoll gradients
Authors: AITKEN R. J. , WEST K. M. 
Published: May 2008

SummaryWe have shown previously that the reactive oxygen species generated by washed human ejaculates originate from cells which can be isolated in the low density region of discontinuous Percoll gradients. In this study we have used a simplified two‐step (40/80%) Percoll gradient to separate human ejaculates (n=109) into two populations of spermatozoa, exhibiting either a high or a low capacity for reactive oxygen species generation. We have then examined the relationships between this activity and other properties of the isolated fractions, with particular emphasis on the presence of leucocytes, which we have quantified using a monoclonal antibody directed against the common leucocyte antigen. The low‐density cells recovered from the 40%/80% interface of the Percoll gradients, differed from the high‐density fraction in exhibiting significantly reduced sperm motility, poorer sperm morphology, and a considerably enhanced capacity for reactive oxygen species production (P<0.001). In six cases the elevated levels of reactive oxygen species generation were associated with leucocyte concentrations in excess of 1 times 106 per 107 sperm, suggesting that leucocytes enter the seminal compartment in an activated, oxygen‐radical generating, state. However, in the majority of cases exhibiting high levels of reactive oxygen species production, leucocyte numbers were low or absent and the semen profiles were unremarkable, except that seminal sperm concentrations tended to be low. These results suggest that the oxidative stress responsible for defective sperm function involves reactive oxygen species originating from two sources; the sperm and infiltrating leucocytes.
Aniline blue staining as a marker of sperm chromatin defects associated with different semen characteristics discriminates between proven fertile and suspected infertile men
Authors: AUGER J. , MESBAH M. , HUBER C. , DADOUNE J. P. 
Published: May 2008

SummaryA retrospective study of 49 men with proven fertility and 396 suspected infertile men was conducted with the primary objective of investigating the relationship between the nuclear maturity of sperm and male fertility. Acidic aniline blue staining was used to detect chromatin defects of sperm nuclei related to their nucleoprotein content as associated with DNA. The discriminant value of the percentage of unstained nuclei (= percentage of mature heads, MH) and of other semen characteristics, was analysed by a stepwise (forward) linear regression model. Semen characteristics that discriminated significantly between the two groups of subjects were, in descending order: (1) the percentage of normal sperm, (2) the percentage of amorphous heads, (3) the percentage of tapered heads, (4) semen volume, and (5) the percentage of MH. The discriminant value of each of the significant characteristics was studied by means of ROC‐curves. MH had the best ROC‐curve profile; its cut‐off value was found to be equal to 70% (74.5 ± 2.6% for the donor group versus 53.0 ± 1.1% for the patient group). A simple infertility score (SIS) including MH, was built according to the cut‐off values inferred from the ROC‐curves. SIS allowed an overall satisfactory separation of the two groups (≤4 = fertile, 5–6 = uncertainty zone, >6 = infertile). Our results indicate that the addition of the evaluation of sperm head maturity to routine semen analysis improves the assessment of fertility in men.
Carcinoma‐in‐situ cells in cultured human seminiferous tubules
Authors: LAUKE H. , SEIDL K. , HARTMANN M. , HOLSTEIN A. F. 
Published: May 2008

SummaryFor the first time, early germ‐cell derived tumour cells were studied in an in‐vitro system of cultured seminiferous tubules. The intratubular tumour cells not only survived in culture for 7 days but were also able to multiply. Dividing tumour cells were identified in semi‐thin sections and electron micrographs by morphological criteria. Additionally, mitotic activity was demonstrated by ‘Hlthymidine histo‐autoradiography’. There are numerous reports on cell lines established from solid non‐seminomas, but up to now no references to seminoma cell lines or cultures of intratubular tumour cells are available. The culture of seminiferous tubules offers a tool in making carcinoma‐in‐situ cells accessible for experimental work.
Effects of seminal plasma from normal and asthenozoospermic men on the progressive motility of washed human sperm
Authors: MAGNUS Ø. , BREKKE I. , ÅBYHOLM T. , PURVIS K. 
Published: May 2008

SummaryHuman seminal plasma stimulated the progressive motility of human sperm in a dose‐ and time‐related manner. Serum exhibited a similar capacity for stimulation. Seminal plasma from different normozoospermic men showed marked variation in its capacity to stimulate sperm motility. The stimulatory effect was maintained after heat‐treatment, indicating the action of a low molecular weight substance or metal ion. No differences could be observed in the capacity for stimulation between seminal plasma from normozoospermic (n = 23) and asthenozoospermic (n = 22) men, when tested at the same dilution and under identical conditions. It is concluded that in general, differences in seminal plasma composition cannot account for the reduced sperm motility in asthenozoospermic men. Furthermore, the stimulatory effects of seminal plasma may be a property shared by other biological fluids.
Variation of semen quality in normal men
Authors: MALLIDIS C. , HOWARD E. J. , BAKER H. W. G. 
Published: May 2008

SummaryTo examine the amount of variability in semen analysis results and whether there is any effect of season, 673 specimens provided by seven normal, healthy men (61–205 specimens/subject) over 72–324 weeks were assessed for sperm concentration, ejaculate volume, motility and motility index. Noticeable sample to sample variations were found. The largest proportion of the overall variance was due to within‐subject differences, e.g., sperm concentration (54%), ejaculate volume (59%), percentage motility (96%) and motility index (74%). Although changes in semen‐analysis results occurred over the year, no consistent trend was seen. No evidence was found to suggest that the differences were due to modifications of the methods employed by the laboratory, or to the change of season.
The effect of A23187 concentration and exposure time on the outcome of the hamster egg penetration test
Authors: FORD W. C. L. , REES J. M. , MCLAUGHLIN E. A. , GODDARD R. J. , HULL M. G. R. 
Published: May 2008

SummaryWe investigated the conditions required to enhance the performance of human sperm in the hamster egg penetration test with the free acid form of A23187. The best performance was observed after stimulation with 2 μM A23187 for 1 h when the median penetration rate with sperm from fertile donors was 100% of eggs with 5.8 decondensed sperm headdegg. Extending the stimulation period with 2 μm A23187 to 2 or 3 h, resulted in a progressive decrease in the penetration rate. In the absence of A23187, the penetration rate was lower (0.7 decondensed sperm heads/ egg after 1 h) but increased with stimulation time. A similar picture was observed with sperm from patients taken for an IVF programme. For a pool of cryopreserved semen, the coefficient of variation of the penetration rate after stimulation with 2 μM A23187 for 1 h, expressed as decondensed sperm headdegg, was 11% within and 20% between assays. There was no correlation between the outcome of the hamster egg penetration test and the percentage motility, velocity or lateral head displacement of the sperm measured after the same stimulation regime. However, in IVF patients the initial velocity and lateral head displacement of the sperm (zero time) were correlated with the best result from the hamster egg penetration test (r= 0.62 and 0.57 respectively). No motility changes characteristic of capacitation were detected. We conclude that stimulation with 2 μM A23187 (free acid) for 1 h prior to the addition of the zona free hamster eggs can produce a high penetration rate with fertile samples and provides a convenient and robust protocol for the assay. However, when carried out in this way the test does not assess the ability of the sperm to capacitate.
Detection of antisperm‐antibodies in patients with primary testicular cancer
Authors: FOSTER R. S. , RUBIN L. R. , MCNULTY A. , BIHRLE R. , DONOHUE J. P. 
Published: May 2008

SummarySerum antisperm‐antibodies have been noted to develop as a result of various types of testicular pathology and injury. Their presence sometimes correlates with impaired fertility. We examined 52 patients with non‐seminomatous low‐stage testicular cancer for the presence of serum antisperm‐antibodies to determine whether or not auto‐immunity is a factor in the depressed fertility known to exist in these patients. Twenty‐one per cent were discovered to have serum antisperm‐antibodies using an immunofluorescent technique.