Studies were conducted to determine the efficiency of decondensation protocols. Sperm obtained from seven normal donors was immediately washed after liquefaction and then decondensed using the method of West or al. (1989) and my original protocol. My optimized protocol entailed mixing 1 ml aliquots of semen with 4 ml phosphate buffered saline (PBS) Following centrifugation, pellets were resuspended in 1 ml PBS containing 6 mM EDTA. After centrifugation, pellets were resuspended in 1 ml PBS containing 2 mM dithiothreitol at 37degrees C for 45 min. Following mixing with 2 ml PBS and centrifugation, pellets were resuspended by vortexing. While vortexing, 5 ml of fixative were gently added. Slide preparation was accomplished using the smear method and it was stored at 4degrees C. When comparing these protocols, the degree of sperm decondensation and head swelling was monitored by measuring nuclear length, area, perimeter, and degree of roundness using FISH analysis software. Apparent copy number for chromosome 1 and, separately, for the sex chromosomes was determined by FISH using satellite DNA probes for loci DIZ1, DXZ1 and DYZ3. Sperm treated by my decondensation protocol showed significant increases (p<0.05) in length, area, perimeter, and degree of roundness. There was a significant decrease (p<0.05) in the frequency of nuclei displaying no signal but no change in the frequency of nuclei with two signals in samples decondensed by my protocol.. My data suggested that decondensation using my original protocol may lower the frequency of cells with spurious 'nullisomy' due to hybridization failure without inducing spurious 'disomy' resulting from increased distances between split signals.
Centrifugation ; Chromosomes, Human, Pair 1 ; Dithiothreitol ; DNA, Satellite ; Edetic Acid ; Fluorescence* ; Head ; Humans* ; In Situ Hybridization* ; Semen ; Sex Chromosomes ; Spermatozoa* ; Tissue Donors

SUMMARY: To investigate the influences on semen parameters and fertilizing capacity of immuno- globulin(Ig) isotypes and regional distribution of antisperm antibody (ASA) on the human sperm surface. Sixty-seven ASA-positive patients were compared with 96 ASA-negative donors. ASAs in semen showed significant negative effects on both semen parameters and fertilizing capacity; in those with ASAs in the sperm head and/or tail, the reductions were significant. In the head as well as the tail, there was close correlation between fertilizing capacity and both IgG and IgA. Both semen parameters and fertilizing capacity are significantly affected by the presence of ASA in semen. In particular, antibodies IgG to sperm head and/or tail, and antibodies IgA to sperm tail appeared to have a highly detrimental effect on fertilizing capacity.
Antibodies* ; Head ; Humans ; Immunoglobulin A ; Immunoglobulin G ; Semen* ; Sperm Head ; Sperm Tail ; Spermatozoa ; Tissue Donors

OBJECTIVES: To assess the fertilizing capacity using sperm penetration assay (SPA) to predict the outcome of the in vitro fertilization-embryo transfer (iVF-ET) outcome. MATERIALS AND METHODS: Semen samples were provided by 129 patients undergoing iVF. We attempted to correlate the extent of sperm penetration under enhanced SPA protocol with the results of fertilization, cleavage, preimplantation embryo development, and pregnancy. RESULTS: Univariate analysis demonstrated a statistically significant correlation between fertilizing capacity and motility, kinetics, fertilization, cleavage and embryo development, and pregnancy rate. By logistic regression analysis, fertilizing capacity was found to be the only variable that was statistically significant with respect to pregnancy rate. Fertilizing capacity, cleavage rate and pregnant rate were significantly higher in pregnant group. However, the fertilization rates was comparable with both group. CONCLUSIONS: Lower fertilizing capacity could denote a poorer prognosis for establishing a pregnancy, even after satisfactory fertilization rate is achieved.
Embryonic Development* ; Female ; Fertilization ; Fertilization in Vitro* ; Humans ; Kinetics ; Logistic Models ; Pregnancy ; Pregnancy Rate ; Pregnancy* ; Prognosis ; Semen ; Sperm-Ovum Interactions ; Spermatozoa*

No abstract available.
Chromatin* ; Humans* ; Population Characteristics* ; Sperm-Ovum Interactions* ; Spermatozoa*

No abstract available.
Fertilization in Vitro* ; Fertilization* ; Humans*

Bisphenol A (BPA) is an endocrine-disrupting chemical that is capable of interfering with the normal function of the endocrine system in the body. Exposure to this chemical from BPA-containing materials and the environment is associated with deleterious health effects, including male reproductive abnormalities. A search of the literature demonstrated that BPA, as a toxicant, directly affects the cellular oxidative stress response machinery. Because of its hormone-like properties, it can also bind with specific receptors in target cells. Therefore, the tissue-specific effects of BPA mostly depend on its endocrine-disrupting capabilities and the expression of those particular receptors in target cells. Although studies have shown the possible mechanisms of BPA action in various cell types, a clear consensus has yet to be established. In this review, we summarize the mechanisms of BPA action in spermatozoa by compiling existing information in the literature.
Consensus ; Endocrine Disruptors ; Endocrine System ; Humans ; Male ; Oxidative Stress ; Spermatozoa

The evaluation of infertility in males consists of physical examination and semen analyses. Standardized semen analyses depend on the descriptive analysis of sperm motility, morphology, and concentration, with a threshold level that must be surpassed to be considered a fertile spermatozoon. Nonetheless, these conventional parameters are not satisfactory for clinicians since 25% of infertility cases worldwide remain unexplained. Therefore, newer tests methods have been established to investigate sperm physiology and functions by monitoring characteristics such as motility, capacitation, the acrosome reaction, reactive oxygen species, sperm DNA damage, chromatin structure, zona pellucida binding, and sperm-oocyte fusion. After the introduction of intracytoplasmic sperm injection technique, sperm maturity, morphology, and aneuploidy conditions have gotten more attention for investigating unexplained male infertility. In the present article, recent advancements in research regarding the utilization of male fertility prediction tests and their role and accuracy are reviewed.
Acrosome Reaction ; Aneuploidy ; Chromatin ; DNA Damage ; Fertility* ; Humans ; Infertility ; Infertility, Male ; Male* ; Physical Examination ; Physiology ; Reactive Oxygen Species ; Semen Analysis ; Sperm Injections, Intracytoplasmic ; Sperm Motility ; Spermatozoa ; Zona Pellucida

No abstract available.
Female ; Humans ; Infertility* ; Pregnancy*

No abstract available.
Animals ; Cricetinae* ; Humans* ; Oocytes*

OBJECTIVE: To evaluate the effects of the reactive oxygen species (ROS) generated with a xanthine(X) and xanthine oxidase (XO) system on sperm function, the change of sperm characteristics, lipid peroxidation, and DNA fragmentation in bovine spermatozoa. MATERIALS AND METHODS: ROS were produced using a combination of 100 micrometer X and 50 mU/ml XO. The ROS scavengers: superoxide dismutase (SOD)(200mu/ml) and catalase (500mu/ml) were also tested. Spermatozoa were incubated for 2 hours in BWW medium with a combination of X-XO supplemented with or without ROS scavengers at 37degrees C under 5% CO2 incubator. Sperm movement characteristics by CASA (computer-aided sperm analysis), HOST (hypoosmotic swelling test), Ca-ionophore induced acrosome reaction, malondialdehyde formation for the analysis of lipid peroxidation, the percentage of DNA fragmentation using the method of TdT-mediated nick end labelling (TUNEL) by flow cytometry were determined after 2 hours incubation. RESULTS: The action of ROS on bovine spermatozoa resulted in a decreased in capacity for sperm motility, Ca-ionophore induced acrosome reaction and membrane integrity, an increased in malondialdehyde formation and the percentage of sperm with DNA fragmentation. In the effects of antioxidant, catalase completely alleviated the toxic effects induced by the ROS in terms of sperm function and characteristics, however SOD exhibited no capacity to reduce the toxic effects. CONCLUSION: The ROS can induce significant damages to sperm functions and characteristics. The useful ROS scavengers can minimized the defects of sperm function and various damages of spermatozoa.
Acrosome Reaction ; Catalase ; DNA Fragmentation* ; DNA* ; Flow Cytometry ; Incubators ; Lipid Peroxidation* ; Malondialdehyde ; Membranes ; Reactive Oxygen Species* ; Sperm Motility ; Spermatozoa* ; Superoxide Dismutase ; Xanthine Oxidase