			Home About us Contact

Semen Samples (semen + sample)

Distribution by Scientific Domains

Medical Sciences	84%
Life Sciences	13%

Selected Abstracts

Ultrastructure of the Spermatozoa of the Yangtze Finless Porpoise (Neophocaena phocaenoides asiaeorientalis)

ANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 4 2009
H. Y. Li
Summary Semen sample was collected from two captive adult Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis) during physical examination. One individual was aged about 9 years with body length 143 cm (total length) and body weight 46.1 kg in 2003. The age of the other was unknown and its body length was 147 cm and body weight was 43 kg in 2004. Ultrastructure of their spermatozoa was examined using scanning and transmission electron microscope. The sperm concentration was 4.17 × 109 spermatozoa per ml by the cytometer. The approximate dimensions of the spermatozoa were as follows: head length, 3.366 ± 0.140 ,m (mean ± SE, n = 15); head width, 1.896 ± 0.099 ,m (n = 15); and neck length, 1.004 ± 0.074 ,m (n = 10). The tail included midpiece, principal piece and terminal piece. The length of the midpiece was 1.882 ± 0.077 ,m (n = 9). There is no apparent boundary between the principal piece and the terminal piece, so the length of the principal piece and the terminal piece was 44.612 ± 3.485 ,m (n = 5). Total length of the spermatozoa was 53.314 ± 4.580 ,m (n = 10). The acrosome covered approximately 45.8% of the anterior portion of the head. [source]

Interaction between leucocytes and human spermatozoa influencing reactive oxygen intermediates release

INTERNATIONAL JOURNAL OF ANDROLOGY, Issue 2 2004
Monika Fr
Summary The relationship between the presence of white blood cells (WBCs) and the fertilizing potential of human semen is still an open question. It is well known that the presence of leucocytes in human semen can be related to the production of reactive oxygen intermediates (ROI). Semen samples were obtained from 15 normozoospermic men and leucocytes were isolated from heparinized blood drawn from 15 volunteers. Lucigenin and luminol-mediated chemiluminescence assays were used to determine reactive oxygen species (ROS) generation by non-activated or activated leucocytes through 12-myristate-13-acetate or N-formyl-methionyl-leucyl-phenyalanine (FMLP) before the addition of spermatozoa isolated by swim-up or Percoll procedures. All spermatozoal fractions used in this study were characterized by defining their motility, morphology and viability. The levels of ROS formation by non-activated as well as stimulated leucocytes were significantly decreased after addition of swim-up separated spermatozoa (p < 0.01). The ability to inhibit the basal chemiluminescence was of lower degree for spermatozoa isolated from 90% Percoll fractions than for swim-up sperm. However, addition of sperm cells from 47% Percoll fraction was found to increase both lucigenin and luminol signals. Moreover, the determined ROI levels changed depending on the type of inducing factor used for oxidative burst. Then, spermatozoa selected by swim-up procedure although with only slightly higher viability and morphology than sperm obtained from 90% Percoll fraction clearly exhibited much higher capacity to inhibit ROI secretion by receptor-stimulated leucocytes (FMLP-activation) than Percoll fractionated sperm. Such results may indicate that within normal semen may exist sperm subpopulations with different biochemical mechanisms controlling the interaction between spermatozoa and contaminating leucocytes. When ROI levels contained in normozoospermic semen are dependent on the WBCs activation, it seems that spermatozoa with preserved normal functional competence are able to defend themselves against leucocytes-derived ROI. Also for normozoospermic ejaculates, swim-up sperm may improve semen antioxidant characteristics when comparing with Percoll (90%) separated sperm. It may help for optimal sperm preparation when assisting to infertility treatment. [source]

Aneuploidy in spermatozoa of infertile men with teratozoospermia

INTERNATIONAL JOURNAL OF ANDROLOGY, Issue 4 2001
Kati HÄrkönen
Recent studies have shown that aneuploidy in spermatozoa of infertile men with poor semen quality is increased. The purpose of this study was to determine whether poor sperm morphology is associated with the incidence of spermatozoa with numerical chromosome abnormalities. Semen samples from 20 infertile teratozoospermic men were studied using multicolour fluorescence in situ hybridization (FISH). Men were divided into four groups according to the proportion of normal sperm morphology: infertile men with <10% (group A, n=7), 10,19% (group B, n=6), and 20,29% (group C, n=7) of morphologically normal spermatozoa, and controls (group D, n=5) with ,30% normal forms. Two hybridizations were performed. All the samples were analysed using probes for chromosomes 1 and 7 and, in addition, in group A and in controls with normal semen parameters probes for chromosomes X, Y and 18 were also used. Ten thousand spermatozoa were scored per hybridization. Severely teratozoospermic men (<10% normal forms) had significantly higher frequency of disomy 7, 18, YY, XY and diploidy in their spermatozoa when compared with controls. The results suggest that poor sperm morphology is associated with numerical chromosome abnormalities of spermatozoa. Severely teratozoospermic men may be at an increased risk of producing aneuploid offspring. [source]

Hypo-osmotic swelling test and unexplained repeat early pregnancy loss

JOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH (ELECTRONIC), Issue 1 2010
Sudhindra M. Bhattacharya
Abstract Aim:, To study the relationship of various sperm characteristics and hypo-osmotic swelling test (HOS test) with repeat unexplained early pregnancy loss. Methods:, Semen samples from husbands of 74 couples with a history of repeat early pregnancy loss (group A) were analyzed according to World Health Organization criteria, and a HOS test was performed in each case. Semen samples from 65 husbands with proven fertility (group B) were also studied for comparison. Results:, No statistically significant differences were noted in the age of the husbands, sperm concentration, sperm morphology and percent motile sperm between groups A and B. The mean HOS test scores of the two groups were significantly different (group A: 60.4%; group B: 76.9%; P = 0.01 [normal value: >60%]). In group A, 33.8% of cases (25/74) and in group B, 12.3% of cases (8/65) showed low HOS test scores. Conclusion:, The sperm HOS test may be helpful to screen for any paternal factor associated with repeat embryonic or early fetal loss and in a resource-poor setting, and may be utilized in any clinical laboratory. [source]

Decreased protein tyrosine phosphorylation and membrane fluidity in spermatozoa from infertile men with varicocele

MOLECULAR REPRODUCTION & DEVELOPMENT, Issue 12 2006
M.G. Buffone
Abstract Varicocele is a prevalent pathology among infertile men. The mechanisms linking this condition to infertility, however, are poorly understood. Our previous work showed a relationship between sperm functional quality and the ability of spermatozoa to respond to capacitating conditions with increased membrane fluidity and protein tyrosine phosphorylation. Given the reported association between varicocele, oxidative stress, and sperm dysfunction, we hypothesized that spermatozoa from infertile patients with varicocele might have a combined defect at the level of membrane fluidity and protein tyrosine phosphorylation. Semen samples from infertile patients with and without grade II/III left varicocele were evaluated for motion parameters (computer-assisted semen analysis [CASA]), hyperactivation (CASA), incidence and intensity of protein tyrosine phosphorylation (phosphotyrosine immunofluorescence and western blotting), and membrane fluidity (Laurdan fluorometry), before and after a capacitating incubation (6 hr at 37°C in Ham's F10/BSA, 5% CO2). Spermatozoa from varicocele samples presented a decreased response to the capacitating challenge, showing significantly lower motility, hyperactivation, incidence and intensity of tyrosine phosphorylation, and membrane fluidity. The findings reported in this article indicate that the sperm dysfunction associated to infertile varicocele coexists with decreased sperm plasma membrane fluidity and tyrosine phosphorylation. These deficiencies represent potential new pathophysiological mechanisms underlying varicocele-related infertility. Mol. Reprod. Dev. 73: 1591,1599, 2006. © 2006 Wiley-Liss, Inc. [source]

Protocol Optimization for Long-Term Liquid Storage of Goat Semen in a Chemically Defined Extender

REPRODUCTION IN DOMESTIC ANIMALS, Issue 6 2009
B-T Zhao
Contents A specific problem in the preservation of goat semen has been the detrimental effect of seminal plasma on the viability of spermatozoa in extenders containing egg yolk or milk. The use of chemically defined extenders will have obvious advantages in liquid storage of buck semen. Our previous study showed that the self-made mZAP extender performed better than commercial extenders, and maintained a sperm motility of 34% for 9 days and a fertilizing potential for successful pregnancies for 7 days. The aim of this study was to extend the viability and fertilizing potential of liquid-stored goat spermatozoa by optimizing procedures for semen processing and storage in the mZAP extender. Semen samples collected from five goat bucks of the Lubei White and Boer breeds were diluted with the extender, cooled and stored at 5°C. Stored semen was evaluated for sperm viability parameters, every 48 h of storage. Data from three ejaculates of different bucks were analysed for each treatment. The percentage data were arcsine-transformed before being analysed with anova and Duncan's multiple comparison test. While cooling at the rate of 0.1,0.25°C/min did not affect sperm viability parameters, doing so at the rate of 0.6°C/min from 30 to 15°C reduced goat sperm motility and membrane integrity. Sperm motility and membrane integrity were significantly higher in semen coated with the extender containing 20% egg yolk than in non-coated semen. Sperm motility, membrane integrity and acrosomal intactness were significantly higher when coated semen was 21-fold diluted than when it was 11- or 51-fold diluted and when extender was renewed at 48-h intervals than when it was not renewed during storage. When goat semen coated with the egg yolk-containing extender was 21-fold diluted, cooled at the rate of 0.07,0.25°C/min, stored at 5°C and the extender renewed every 48 h, a sperm motility of 48% was maintained for 13 days, and an in vitro -fertilizing potential similar to that of fresh semen was maintained for 11 days. [source]

Effects of Oxygen Exposure and Gentamicin on Stallion Semen Stored at 5 and 15°C

REPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2008
S Price
Contents This study was undertaken to investigate the effects of storage of stallion semen in a defined milk protein extender at 5 and 15°C under either anaerobic or aerobic conditions, with or without addition of the antibiotic gentamicin. Semen samples were collected from eight fertile stallions and stored for 96 h (day 0,4) and assessed daily for motility, velocity and membrane integrity (viability) using a CASA system. Samples for bacteriology assessment were taken on day 2 of storage. No significant (p > 0.05) differences in motility, velocity or viability were observed between treatments on days 0,2. On days 3 and 4, semen stored without gentamicin at 5°C had a significantly (p < 0.05) better semen quality compared with storage at 15°C without gentamicin, irrespective of oxygen exposure. On days 3 and 4, motility and velocity were greater in samples stored at 15°C with gentamicin, compared with the corresponding treatments without antibiotic (p < 0.05). This effect was also evident for viability on day 4. The decline in semen quality observed at 15°C most likely resulted from the effect of bacterial growth. Bacterial growth was the greatest in samples stored at 15°C without gentamicin, under both anaerobic and aerobic conditions (p < 0.05). Bacterial growth was inhibited by adding of gentamicin at 15°C, which accordingly reduced the decline in semen quality. Addition of antibiotic to samples stored at 5°C had no significant effect on any parameter analysed. In conclusion, storage at 15°C can be achieved by using an extender containing the antibiotic gentamicin. Storage at 5°C tended to maintain better semen quality irrespective of oxygen exposure, and did not necessitate an antibiotic treatment. [source]

Post-thaw Survival and Longevity of Bull Spermatozoa Frozen with an Egg Yolk-based or Two Egg Yolk-free Extenders after an Equilibration Period of 18 h

REPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2007
R Muiño
Contents The aim of the present study was to determine the suitability of using two egg yolk-free commercial extenders, Andromed® and Biociphos Plus®, as compared with the Tris-egg yolk based diluent Biladyl®, for the cryopreservation of bull spermatozoa when the freezing protocol involved holding the extended semen at 4°C for 18 h before the freezing. Six ejaculates from each of 10 Holstein bulls were collected by using artificial vagina. The ejaculates were evaluated for volume, sperm concentration and motility, divided in to three equal volumes, and diluted, respectively, with the three extenders as specified above. Extended semen was equilibrated for 18 h at 4°C and frozen in 0.25-ml straws. After thawing, 100- ,l aliquots of semen were labelled with SYBR-14, PI and PE-PNA (Phycoerythrin-conjugated Peanut agglutinin) and analysed by flow cytometry at 0, 3, 6 and 9 h after incubation at 37°C. A General Lineal Model procedure for repeated measures was used to determine the effects of extender, bull, replicate and the interaction between them, on sperm viability and acrosomal integrity. Semen samples frozen with Biladyl® showed higher (p < 0.001) sperm survival after 0 h (47.9%) and 9 h (30.3%) of incubation than those frozen with Andromed® (38.5% and 17.3%, after 0 and 9 h respectively) or Biociphos Plus® (34.9% and 21.6%, after 0 and 9 h respectively). The bull and replicate had significant effects (p < 0.001) on both sperm viability and acrosomal integrity, but the interactions between bull and extender and between replicate and extender were not significant. It was concluded that, when holding the semen overnight before freezing, the use of Biladyl® results in higher sperm survival and longevity than the use of Andromed® or Biociphos Plus®. [source]

ORIGINAL ARTICLE: Expression of IL-6, IL-8, TNF-,, IL-10, HSP-60, Anti-HSP-60 Antibodies, and Anti-sperm Antibodies, in Semen of Men with Leukocytes and/or Bacteria

AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 3 2010
Elizabeth Martínez-Prado
Citation Martínez-Prado E, Bermúdez MIC. Expression of IL-6, IL-8, TNF-,, IL-10, HSP-60, anti-HSP-60 antibodies, and anti-sperm antibodies, in semen of men with leukocytes and/or bacteria. Am J Reprod Immunol 2010; 63: 233,243 Problem, Different cellular and biochemical markers have been proposed as indicators of infection-inflammation of male genital tract. Method of study, Semen samples from 80 men attending an andrologic clinic were evaluated to determine the presence of leukocyte, bacteria, antibodies against Chlamydia trachomatis, levels of IL-6, IL-8, IL-10, and TNF-,, HSP-60, anti-HSP-60 antibodies, and anti-sperm antibodies. Results, Leukocytes in semen significantly correlated with an increase in IL-6, IL-8, and TNF-,. The simultaneous presence of pathogens and leukocytes was associated with high levels of IL-8 and TNF-,, whereas IL-6 was more associated with the presence of leukocytes. Anti-HSP-60 antibodies positively correlated with IL-6 and IL-8. The presence of anti-sperm antibodies highly associated with an increase in anti-HSP-60 antibodies. Conclusions, The type of cytokines present in the semen will depend on the single or simultaneous presence of leukocytes and/or pathogens. Chronic male genital tract infections could be associated with the development of anti-HSP-60 antibodies and anti-sperm antibodies. [source]

Increased IL-18 Levels in Seminal Plasma of Infertile Men with Genital Tract Infections

AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 6 2006
Ioannis M. Matalliotakis
Problem Interleukin (IL)-18 is a novel cytokine, previously known as interferon (IFN)- , inducing factor. We evaluated the levels of IL-18 and IFN- , in seminal plasma (SP) of fertile and infertile men. Method of study Semen samples were obtained by masturbation from 80 men, and were examined for the levels of IL-18 and IFN- , by enzyme-linked immunosorbent assay. Seven groups were included: (i) fertile men (n = 18), (i) infertile men with genital tract infections (n = 17), (iii) with varicocele (n = 15), (iv) with Klinefelter syndrome (n = 6), (v) with cryptorchidism (n = 7), (vi) with mumps orchitis (n = 7), and (vii) with idiopathic testicular lesions (n = 10). Results Mean levels of IL-18 were higher in SP from infertile men with genital tract infections compared with SP from other groups except Klinefelter syndrome (P < 0.05). However, no significant differences could be detected for IFN- ,. A significant positive correlations was found between IL-18 and IFN- , in total patient population (P < 0.001). Moreover, a negative correlation was observed between IL-18 and sperm concentrations, and motility (P < 0.01 and <0.03, respectively). Furthermore, there was a positive and statistically significant association between IL-18 and IFN- , levels in SP of infertile men with genital tract infections (P < 0.0001). However, there was no relationship between IL-18 and IFN- ,, and semen parameters in the same group. Conclusion SP IL-18 levels were increased in men with urogenital infections. Thus, the elevated expression of IL-18 in SP may be used as a diagnostic marker in the male genital tract infections. [source]

Distinct Expression Pattern of Cytokines in Semen of Men with Genital Infection and Oligo-Terato-Asthenozoospermia

AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 3 2002
Ioannis Matalliotakis
PROBLEM: The objective of this study was to evaluate the possible relevance of cytokines in seminal plasma (SP) of patients with accessory gland infection and oligo-terato-asthenozoospermia. METHOD OF STUDY: Semen samples were obtained by masturbation from 90 men and were examined for the presence of interleukin (IL)-2, IL-6, IL-8, IL-11 and soluble CD23 (sCD23) by enzyme-linked immunosorbent assay. Five groups were included: (1) fertile men (n=20), (2) infertile men with varicocele and oligo-terato-asthenozoospermia (V-OTA, n=20), (3) infertile men with genital infection and OTA (INF-OTA, n=20), (4) infertile men with idiopathic testicular lesion and OTA (ITL-OTA, n=20) and (5) infertile men with azoospermia (AZOO, n=10). RESULTS: We found that the mean level of IL-2 was higher in SP from infertile men compared with SP from fertile men (P < 0.05). Mean levels of IL-6, IL-8, IL-11 in SP of INF-OTA were higher than that of all other groups (P < 0.05, P < 0.05, P < 0.001, respectively). However, no significant differences could be detected between other groups. A significant increase was noted in sCD23 levels in SP from men with ITL-OTA compared with all other groups (P < 0.01). We have not observed any correlations between IL-2, IL-6, IL-8, IL-11 and sCD23 levels in SP and semen parameters. Spearman's correlation coefficient revealed that there was a significant association between IL-6, IL-8, IL-11 levels in men with INF-OTA. CONCLUSION: The measurement of each cytokine separately in the SP of men with INF-OTA, in spite of the existing significant differences, does not have a diagnostic value in male infertility. However, a combined determination of IL-6, IL-8, IL-11 in the SP of men with genital infection and oligo-terato-asthenozoospermia may provide clinically useful information for the diagnosis of male accessory gland infection. [source]

Effects of varicocele upon the expression of apoptosis-related proteins

ANDROLOGIA, Issue 4 2010
F.-W. Chang
Summary Varicocele-associated apoptosis has been recognised as a cause of male infertility. Thus, we assessed the expression of somatic apoptosis-related proteins (the typical protein-dependent apoptosis markers) in ejaculated sperm plasma from both patients with varicocele and normal donors. We evaluated the relationships between certain apoptosis-related proteins and normal semen quality. Semen samples were obtained from 25 patients with varicocele and from 10 normal fertile controls. These samples were compared using computer-assisted semen analysis for motion parameters and manual analysis for morphology, and were also assayed for apoptosis-related protein activation including caspase-3, poly-ACP-ribose polymerase (PARP), the Bcl-2 family (Bcl-2, Bak) and p53 by means of immunoblot analysis. PARP, Bak and p53 were expressed substantially more in the sperm cells of the varicocele group when compared with the normal group (P < 0.05). The expression of caspase-3 and Bcl-2 did not appear to differ between these two study groups. An increased expression of PARP, Bak and p53 for varicocele-afflicted individuals indicated an increased participation by these agents in the regulating of apoptosis in the ejaculated semen from patients with varicocele, suggesting that certain protein-development apoptotic mechanisms might originate in the cytoplasmic droplet or within mitochondria of spermatocytes and then might function within the nucleus of the cell. [source]

CASA-based sperm kinematics of environmental risk factor-exposed human semen samples designated as normozoospermic in conventional analysis

ANDROLOGIA, Issue 4 2010
D. Mukhopadhyay
Summary This study was conducted after an initial epidemiological survey of patients in and around Calcutta, India, concerning their lifestyle history, degree of risk exposure and semen analysis based on conventional WHO criteria. It was found that a large group of exposed patients were showing normozoospermic semen parameters in conventional semen analysis. Hence, a selected group of subjects, designated as normozoospermic in routine analysis, but under risk factor exposure, were selected for a repeat computer aided semen analysis (CASA) and were compared with a control group. The parameters considered among CASA results were: curvilinear velocity (VCL), straight-line velocity, average path velocity (VAP), straightness index (STR), lateral head displacement (ALH) and beat cross frequency. The results depict a significant decline in the mean values of VCL (P = 0.029) and STR (P = 0.007) in the tobacco-exposed group when compared with the unexposed group. On the other hand, there was a significant decline in the mean values of VCL (P = 0.014) and ALH (P = 0.040) in the heavy metal-exposed group when compared with the unexposed group. The other parameters did not show significant change in either group. Semen samples that had been designated normozoospermic in conventional analysis were seen to be influenced by risk factors at the level of sperm motion kinetics. [source]

Impact of chronic viral diseases on semen parameters

ANDROLOGIA, Issue 2 2010
F. Lorusso
Summary The aim of this study was to assess the effect of human immunodeficiency virus (HIV), hepatitis C (HCV) and B (HBV) virus infection on semen parameters. Semen samples were obtained from 27 HCV, 34 HIV, 30 HBV and 41 HCV-HIV-seropositive patients and compared with those of a control population of healthy seronegative subjects. Tests for detection of HIV, HCV and HBV were performed on seminal samples. The sperm concentration was significantly decreased in HCV- and HBV-seropositive males compared to that of controls (P < 0.001). The mean sperm motility (a + b) was significantly decreased in HCV- and HBV-seropositive (P < 0.001) and in HCV-HIV-seropositive subjects (P < 0.05) compared to that of controls. The sperm viability was significantly lower in HCV- and HBV-seropositive men than in controls (P < 0.001). The normal morphology was significantly reduced in HCV-seropositive and HBV-seropositive men (P < 0.05) with respect to that of controls (P < 0.05). The sperm concentration after sperm wash was significantly higher in controls than in HCV-, HIV-, HBV- and HIV-HCV-seropositive men (P < 0.001). We can conclude that HBV- and HCV-infected men have a significantly impaired sperm quality compared with that of controls. The reason for the better sperm quality in our series of HIV- and HCV-HIV-infected men is still under debate. Further investigations in a larger case series are warranted. [source]

Effects of post-density gradient swim-up on apoptosis signalling in human spermatozoa

ANDROLOGIA, Issue 2 2010
S. Grunewald
Summary The inclusion of apoptotic spermatozoa during assisted reproductive techniques (ART) may be one reason for suboptimal success rates. The aim of our study was to evaluate the potential of routine semen preparation to eliminate spermatozoa with activated apoptosis signalling. Semen samples from 20 infertility patients scheduled for ART procedures were investigated. Following density gradient centrifugation (DGC) and swim-up, aliquots were taken from each sample to analyse motility, Caspase-3 activation (CP3) and integrity of the mitochondrial membrane potential (MMP) using flow cytometry. Aliquots from the neat semen served as controls. Semen samples of patients contained 53.8 ± 17.7% spermatozoa with disrupted MMP and 51.8 ± 14.9% with active CP3. Preparation by DGC and swim-up resulted in improvement of progressive motility (+43.5%) and reduction of spermatozoa with disrupted MMP (,34.3%) and activated CP3 (,25.7%, P < 0.01). Minimal reduction of spermatozoa with disrupted MMP and active CP3 was 6.0% and 0.7%, maximum reduction was 65.5% (disrupted MMP) and 49.3% (CP3). Semen samples of subfertile patients contain high levels of spermatozoa with activated apoptosis signalling. Although there was a reduction in the majority of the samples, profound interindividual differences in the separation effect demand further development of innovative molecular-based separation methods to deplete apoptotic spermatozoa. [source]

Evaluation of zeta and HA-binding methods for selection of spermatozoa with normal morphology, protamine content and DNA integrity

ANDROLOGIA, Issue 1 2010
S. H. Razavi
Summary Sperm selection parameters based on morphology and motility for ICSI might not be relevant to chromatin integrity. Thus sperm selection based on sperm characteristics has been suggested. Therefore, the aim of this study was to evaluate the efficiency of the zeta and hyaluronic acid (HA) sperm selection procedures with neat semen, for recovering spermatozoa with normal morphology, protamine content and DNA integrity in infertile men. Semen samples from 77 infertile couples were assessed during this study. Semen analysis was carried out according to World Health Organization criteria. Protamine content, DNA integrity and sperm morphology were assessed by chromomycin A3, sperm chromatin dispersion and Papanicolaou staining respectively. The results show that both HA and zeta methods were efficient to recover spermatozoa with normal morphology and protamine content. In terms of the latter parameters, there was no superiority between the two procedures. However, in terms of DNA integrity, the zeta method was more efficient compared with the control and HA procedure and no significant difference was observed between HA and the controls. Therefore, the zeta method appears to be a suitable procedure to recover spermatozoa with normal DNA integrity. [source]

Predictors of improved seminal parameters and fertility after varicocele repair in young adults

ANDROLOGIA, Issue 5 2009
M. Rodriguez Peña
Summary The aim of our study was to determine hormonal or biochemical markers in patients with clinically palpable left varicocele but without a history of infertility, with especial emphasis on nitric oxide, related with improved seminal parameters after varicocelectomy. Semen samples were obtained from 202 patients with left varicocele grade II or III. Nitric oxide levels in seminal plasma were determined by the Griess technique. Testicular volume was determined ultrasonographically in both testes and hormonal profile was measured. The post-operative sperm concentration increased significantly in patients with normal sperm count or moderate oligozoospermia, but we did not find an increment in sperm count in patients with mild and severe oligozoospermia after surgery. The mean percentage of normal motility significantly increased after surgery, but we did not observe a significant increment in morphologically normal sperm count and testicular volume after varicocele repair. Moreover, we did not find any correlation between nitric oxide concentrations and severity of oligozoospermia, asthenozoospermia or abnormal sperm morphology in this population. It is concluded that in the general male population, varicocele repair is not associated with an improved semen profile in all cases. We did not observe a significant correlation between nitric oxide concentrations and semen profile. [source]

Seminal plasma albumin: origin and relation to the male reproductive parameters

ANDROLOGIA, Issue 2 2007
S. Elzanaty
Summary We wanted to investigate the origin of seminal plasma albumin and its relation to the male reproductive parameters. Semen samples from 916 men, under infertility assessment, were analysed according to guidelines of the World Health Organization. Seminal plasma constituents, i.e. albumin, markers of the epididymal (neutral , -glucosidase, NAG), prostatic (prostate-specific antigen, PSA, and zinc) and seminal vesicle function (fructose), as well as levels of reproductive hormones in plasma were measured. The sperm chromatin structure assay (SCSA) was applied on 267 of the 916 samples. A negative correlation was seen for seminal albumin and plasma follicle-stimulating hormone (r = ,0.1, P = 0.02) and a positive correlation for seminal albumin and serum inhibin B (r = 0.2, P = 0.004). Albumin exhibited positive correlations with the epididymal marker, NAG (r = 0.5, P < 0.001) and with the prostatic markers, PSA and zinc (r = 0.1, P = 0.001; r = 0.2, P < 0.001 respectively) as well as with age (r = 0.2, P < 0.001). A negative significant association was seen for seminal albumin and semen volume (, = ,0.60; 95% CI ,0.80 to ,0.30). The opposite trend was found regarding sperm concentration (, = 0.34; 95% CI 0.30,0.40), total sperm count (, = 0.30; 95% CI 0.20,0.40), and percentage morphologically normal spermatozoa (, = 0.70; 95% CI 0.10,1.0). No association was found between albumin and sperm motility, SCSA parameters, or fructose, the marker of seminal vesicles. Our results suggest testicular, epididymal and prostatic origin of seminal plasma albumin, in addition to the contribution from blood. This is the first study to demonstrate an association between seminal plasma albumin and sperm morphology. Further studies are needed to elucidate the role of seminal albumin in sperm morphology. [source]

Tadalafil as an in vitro sperm motility stimulant

ANDROLOGIA, Issue 1 2007
T. Mostafa
Summary Tadalafil (Cialis®) is a known oral selective phosphodiesterase-5 inhibitor used widely in the management of erectile dysfunction. To assess its ability on human sperm motility in vitro, 70 asthenozoospermic semen specimens delivered by masturbation were investigated. Semen samples were divided equally into four tubes, one as a control and to the others tadalafil dissolved solution was added in vitro in three different concentrations (4.0, 1.0, 0.5 mg ml,1 respectively). The tubes were incubated and were followed up for sperm motility per cent changes for 0.5, 1, 2, 3 h. It was found that the concentration used played an important role in the degree of sperm enhancement. Specimens treated with 4 mg ml,1 tadalafil solution demonstrated a significant decrease in sperm motility compared with the controls. Specimens treated with 1.0 mg ml,1 solution demonstrated significant increase in sperm progressive forward motility. Specimens treated with 0.5 mg ml,1 solution demonstrated significant increases in sperm motility but lower than that of 1 mg ml,1 concentration. It is concluded that in vitro use of tadalafil solution in special concentration has a significant stimulatory effect on asthenozoospermic sperm motility. [source]

TEM, FISH and molecular studies in infertile men with pericentric inversion of chromosome 9

ANDROLOGIA, Issue 4 2006
G. Collodel
Summary Pericentric inversions involving the secondary constriction (qh) region of chromosome 9 are considered to be normal variants of human karyotype. A number of investigators have suggested that chromosomal anomalies can contribute to human infertility causing spermatogenetic derangement. The present study was aimed at verifying the influence of chromosome 9 inversion on human spermatogenesis. Semen samples of 18 male carriers of chromosome 9 inversion, analysed by light microscopy, revealed that five patients were azoospermic. PCR analysis demonstrated that two of them also had Y microdeletions. The other 13 showed generally normal sperm concentrations and reduced motility. The morphological characteristics of sperm were studied by TEM and the data were elaborated by a mathematical formula. Sperm pathologies resulted more frequently in the studied group compared to controls, particularly apoptosis. Partial sequences of the A-kinase anchoring protein (Akap) 4 and 3 genes were performed in all patients, as a previous study by our group highlighted Dysplasia of Fibrous Sheath (DFS) defect in two men with inv 9 investigations. The possible effect of chromosome 9 inversion on meiotic chromosome segregation was investigated by FISH, which showed an increased incidence of diploidy. We hypothesized that this inversion could have variable effects on spermatogenesis, from azoospermia to severely altered sperm morphology, motility and meiotic segregation. [source]

The limit of leucocytospermia from the microbiological viewpoint

ANDROLOGIA, Issue 5 2003
Margus Punab
Summary. The aim of the study was to find out the correlation between white blood cell (WBC) counts in semen and quantitative composition of seminal microflora, and to establish the minimum WBC count associated with significant bacteriospermia. The research included 159 men with different WBC counts in their semen, 84 of them with chronic prostatitis/chronic pelvic pain syndrome. Semen samples were cultivated quantitatively for detecting anaerobic, microaerophilic and aerobic bacteria. Bryan-Leishman stained slides were used for detecting WBC in semen. Seminal fluid was colonized by eight different microorganisms, and the total count of microorganisms in semen ranged from 102 to 107 CFU ml,1. A high frequency of anaerobic microorganisms was found. A positive correlation was observed between the WBC count and the number of different microorganisms, and also between the WBC count and the total count of microorganisms in semen sample. The receiver operating characteristic curve analysis demonstrated that the WHO-defined WBC cut-off point (1 × 106 WBC ml,1) has very low sensitivity for discriminating between patients with and without significant bacteriospermia, as a more optimal sensitivity/specificity ratio appears at 0.2 × 106 WBC ml,1 of semen. The quantitative microbiological finding of semen in the patients of National Institute of Health (NIH) categories IIIa and IV was very similar, i.e. a high number of different microorganisms and a high total count of microorganisms. In the control group (without leucocytospermia and prostatitis symptoms) both parameters were significantly lower. [source]

Influence of sexual stimulation on sperm parameters in semen samples collected via masturbation from normozoospermic men or cryptozoospermic men participating in an assisted reproduction programme

ANDROLOGIA, Issue 3 2000
Y. Yamamoto
Summary. To evaluate the influence of sexual stimulation via sexually stimulating videotaped visual images (VIM) on sperm function, two semen samples were collected from each of 19 normozoospermic men via masturbation with VIM. Two additional samples were collected from each man via masturbation without VIM. The volume of seminal plasma, total sperm count, sperm motility, percentage of morphologically normal spermatozoa, outcome of hypo-osmotic swelling test and zona-free hamster oocyte sperm penetration assay, and markers of the secretory function of prostate were significantly larger in semen samples collected via masturbation with VIM than masturbation without VIM. The improved sperm parameters in the samples collected via masturbation with VIM may reflect an enhanced prostatic secretory function and increased loading of the vas deferens at that time. In a similar protocol, two semen samples were collected via masturbation with VIM from each of 22 non-obstructed azoospermic men. Semen samples from these men had been occasionally positive in the past for a very small number of spermatozoa (cryptozoospermic men). Two additional samples were collected from each cryptozoospermic man via masturbation without VIM. The volume of seminal plasma, total sperm count, sperm motility, and a marker of the secretory function of prostate were significantly larger in semen samples collected via masturbation with VIM. Fourteen out of the 22 men were negative for spermatozoa in both samples collected via masturbation without VIM. These men demonstrated spermatozoa in both samples collected via masturbation with VIM. Six men with immotile spermatozoa in both samples collected via masturbation without VIM exposed motile spermatozoa in both samples collected via masturbation with VIM. High sexual stimulation during masturbation with VIM results in recovery of spermatozoa of greater fertilizing potential both in normozoospermic and cryptozoospermic men. The appearance of spermatozoa after masturbation with VIM in the vast majority of cryptozoospermic men is of clinical significance in programmes applying intracytoplasmic sperm injections for the management of severe male infertility and obviates the need for testicular biopsy. [source]

Semen quality and sedentary work position

INTERNATIONAL JOURNAL OF ANDROLOGY, Issue 1 2004
Julie Støy
Summary Increased scrotal temperature can, in experimental settings, markedly disturb the production of semen. Sedentary work position may increase the temperature of the scrotum, but previous studies have failed to determine whether changes in scrotal temperature caused by sedentary work actually do affect semen quality. This study was carried out to elucidate the possible harmful effects of sedentary work on sperm count and other semen characteristics. In 1981,1983 a semen sample was obtained from 3119 men who attended an infertility workup in one of four Danish fertility centres. A total of 2517 men returned a postal questionnaire with information on life style, leisure time activities, occupational history and job duties. Information on job specific work position was obtained from The Danish Work Environment Cohort study 1990 (DWECS). In this analysis DWECS data for a total of 1747 men was included from men aged 18,39 years with >30 h of work per week. For all job titles represented in the DWECS, the mean proportion of sedentary work was estimated. The sperm cell concentration was 30.6 million/mL among men in the quintile with lowest job specific sedentary work compared with 40.5 million/mL in the highest quintile. The difference was, however, not statistically significant. Stratification on infertility period, educational level of the man, fertility centre, and fertility-related disease of the spouse did not influence the results. The analyses do not suggest that sedentary work is a risk factor for abnormal semen characteristics. [source]

Comparison between computerized slow-stage and static liquid nitrogen vapour freezing methods with respect to the deleterious effect on chromatin and morphology of spermatozoa from fertile and subfertile men

INTERNATIONAL JOURNAL OF ANDROLOGY, Issue 2 2001
M. E. Hammadeh
The purpose of this study was to determine the negative effects (cryodamage) on human spermatozoa after freeze-thawing and to determine whether freeze-thawing of spermatozoa with a programmed slow freezer is better than freezing with liquid nitrogen vapour (rapid freezing) with regard to alterations in sperm chromatin and morphology in semen from fertile (donor) and subfertile, IVF/ICSI, patients. Ninety-five semen samples were obtained either from patients attending our IVF unit for treatment (n=34) or from donors (n=25) with proven fertility and normal sperm quality according to WHO guidelines. Each semen sample was divided into two parts after liquefaction and addition of the cryoprotectant. The first part was frozen using a programmed biological freezer and the second part was frozen by means of liquid nitrogen vapour. Smears were made before the freezing and after the thawing procedure to assess morphology (strict criteria) and chromatin condensation (Acridine Orange test). The mean percentage of chromatin condensed spermatozoa in the samples from donors (control group) was 92.4 ± 8.4% before freezing and this decreased significantly (p < 0.0001) to 88.7 ± 11.2% after freeze-thawing with the computerized slow-stage freezer and to 87.2 ± 12.3% after using static liquid nitrogen vapour (p < 0.001). The corresponding values for semen obtained from patients was 78.9 ± 10.3% before freezing which decreased to 70.7 ± 10.8 and 68.5 ± 14.8%, respectively (p < 0.001). On the other hand, the mean percentage of normal sperm morphology in the control group decreased from 26.3 ± 7.5% before freezing to 22.1 ± 6.4% (p < 0.0001) after thawing with the computerized slow-stage freezer and to 22.2 ± 6.6% (p < 0.0001) after the use of static liquid nitrogen vapour. In the patient group, the mean percentage of normal morphology decreased from 11.7 ± 6.1% after freezing with the biological freezer to 9.3 ± 5.6% and to 8.0 ± 4.9% after freezing with static liquid nitrogen vapour. This study demonstrates that chromatin packaging and morphology of human spermatozoa decrease significantly after the freeze-thawing procedure, not only after the use of static liquid nitrogen vapour but also after the use of a computerized slow-stage freezer. However, the chromatin of semen samples with normal semen parameters (donor sperm) withstand the freeze-thaw injury better than those with low quality semen samples. Therefore, the computerized slow stage freezer could be recommended for freezing of human spermatozoa, especially for subnormal semen samples, for example, ICSI and ICSI/TESE candidates and from patients with testicular tumours or Hodgkin's disease, in order to avoid further damage to the sperm chromatin structure. [source]

Multivariate Cluster Analysis Regression Procedures as Tools to Identify Motile Sperm Subpopulations in Rabbit Semen and to Predict Semen Fertility and Litter Size

REPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2007
A Quintero-Moreno
Contents Computerized motility analysis (CASA) shows that four separate subpopulations of spermatozoa with different motility characteristics co-exist in rabbit ejaculates. There were significant (p < 0.01) differences in the distribution of these subpopulations among separate genetic lines, total sperm abnormalities and the percentage of altered acrosomes. Furthermore, logistic and linear multivariate regressions among several parameters of rabbit semen quality analysis were tested for use as predictive tools for the fertilizing ability of a specific artificial insemination semen sample. Logistic regression analysis rendered two mathematical, significant (p < 0.01) models: one between sperm viability and conception rate and the other between total sperm abnormalities and conception rate. Multiple linear regression analyses also yielded some significant relationships between both fertility (p < 0.001) and litter size (p < 0.05), with respect to some semen characteristics. Our results support the hypothesis that the predictive in vivo fertility use of the standard rabbit semen quality analysis coupled with a CASA determination could be reasonably achieved by applying linear and logistic regression analyses among several parameters of rabbit semen quality analysis. [source]

Can We Increase the Estimative Value of Semen Assessment?*

REPRODUCTION IN DOMESTIC ANIMALS, Issue 2006
H Rodríguez-Martínez
Contents Estimating the fertility of a semen sample or of the male from where it has been collected by simple assessment of in vitro sperm characteristics is still difficult, owing to the variable correlations that laboratory results achieve with in vivo fertility. A major reason behind these variations is the fact that the ejaculate and the artificial insemination (AI)-doses it generates are composed of a diverse sperm population. Such heterogeneity is reflected both in differences of intactness of attributes needed for fertilization, such as motility or morphology, but also in the relative ability of spermatozoa to prevail fertile over time, handling and exposure to different stimuli, all of which account for innate variations in fertilizing ability among doses, ejaculates and sires. However, methods are already available to select sub-populations of intact spermatozoa which can be tested for their degree of competence for fertilization and whose estimative power is promising, allowing the elimination of cases of sub-fertility, particularly in bovine. Examples of these methods are the separation of viable spermatozoa by swim-up or discontinuous gradient centrifugation, followed by testing the ability of the selected spermatozoa to dose-response/time sustain capacitation and acrosome reaction induction. Finding how large a sperm population with non-compensable attributes for fertilization and ability to display and sustain stimuli is, perhaps by a quick screening of membrane integrity and stability by multi-parametric methods, would allow, provided the particular male produces this sub-population in a repeatable manner, for a better estimation of fertility. [source]

The limit of leucocytospermia from the microbiological viewpoint

Association between freezing agent and acrosome damage of human spermatozoa from subnormal and normal semen

ANDROLOGIA, Issue 6 2001
M. E. Hammadeh
Summary. This experimental study compares the effects of human sperm preservation medium (HSPM) with TEST,yolk buffer (TYB) as cryoprotectants of human spermatozoa with respect to the integrity of the acrosome after the freeze,thawing procedure. Fifty-six semen samples were included in this study; 18 were subnormal (G1) and 38 were normal (G2) based on World Health Organization criteria, except for morphology, which was evaluated according to strict criteria. Each semen sample was divided into two parts: the first part was prepared for cryopreservation by the addition of HSPM (1:1) and the second by addition of TYB (1:1). Freezing was performed in liquid nitrogen vapour. Smears were made before freezing and after the thawing process for evaluation of acrosome integrity using fluorescent-lectin labelling. The mean percentage of spermatozoa with intact acrosomes in the subnormal group was 77.0 ± 7.2% before freezing and decreased significantly (P < 0.001) after thawing: to 63.7 ± 8.2% with the use of HSPM and 66.8 ± 8.7% with the use of TYB. The corresponding values in the normal semen samples were 83.4 ± 9.2%, 76.0 ± 8.8% and 77.9 ± 9.2%, respectively. It is obvious that the decrease in the mean percentage of spermatozoa with intact acrosome was significantly higher when using HSPM in comparison with TYB, not only for G1 (,14.9 ± 1.9% versus ,11.8 ± 1.4%) but also for G2 samples (,13.8 ± 1.5% versus ,11.9 ± 1.3%). In conclusion, TYB should be recommended for freeze,thawing of human spermatozoa as the first-choice cryoprotectant, for normal as well as subnormal semen samples, in order to protect the sperm acrosome from the deleterious effects of the freeze,thawing procedure. [source]

Intravasal azoospermia: a surgical dilemma

BJU INTERNATIONAL, Issue 9 2000
Y.R. Sheynkin
Objectives,To determine the incidence of intravasal azoospermia (IVA) and evaluate which factors before and during surgery influence outcome, by prospectively and intentionally performing bilateral vasovasostomies (VVs) only in men with intraoperative IVA. Patients and methods,Using a multilayer technique, 472 men underwent microsurgical reconstructive procedures. Intravasal fluid was examined for sperm by the surgeon and a pathologist. Strict enrolment criteria included total absence of sperm or sperm parts and bilateral VV as a treatment procedure. Patients were followed up by semen analysis and paternity assessed only by naturally conceived pregnancies. Results,Of the 472 patients, 27 (5.7%) had bilateral IVA; 15 of these patients were available for a follow-up of 1,47 months. Eleven patients had identical gross appearance of intravasal fluid bilaterally. Of these patients, five had sperm in the ejaculate after surgery (three with clear intravasal fluid and two with no fluid). Bilaterally different vasal fluid was found in four men. Unilateral clear fluid was present in three patients, two of whom had sperm in semen analysed after VV. Overall, there was sperm in the ejaculate in seven of 15 patients with IVA; five of these seven had clear fluid in at least one vas deferens. One patient with unilaterally clear fluid achieved paternity by a naturally conceived pregnancy. The difference between the mean (sem) obstruction interval in men who had sperm in a semen sample after VV, at 16.7 (3.30) years, and in persistently azoospermic patients, at 15.5 (1.89) years, was not statistically significant (P = 0.741). Conclusion,The results of VV in patients with IVA are unsatisfactory; the patency rate is higher in men with copious clear fluid in at least one vas. The obstructive interval in patients with IVA does not appear to influence the outcome of VV. [source]

Cryopreservation of semen from a stallion with seminal vesiculitis

EQUINE VETERINARY EDUCATION, Issue 5 2010
L. C. Fennell
Summary A 6-year-old Warmblood stallion was admitted for semen collection and cryopreservation. On the seventh and subsequent collection days semen samples were contaminated with purulent debris. A diagnosis of seminal vesiculitis was made following ultrasonography and endoscopy of the seminal vesicles. The stallion was treated with systemic and topical antimicrobial therapy and, although this did not cure the condition, subsequent ejaculates were suitable for cryopreservation. [source]