1,720,985 research outputs found
"Metodiche di valutazione del liquido seminale nella diagnosi dell'infertilità di coppia."
No full textA quick molecular method for the simultaneous detection in spermatozoa of nuclear, acrosomal and axonemal structure by fluorescent microscopy
No full textEjaculated spermatozoa from infertile men presenting to our laboratory for semen analysis were processed with a new molecular method which reveals simultaneously, in the same sperm cell, the status of the acrosome, by testing the hyaluronidase content, the texture of the nucleus, by checking the DNA strands breaks, and the structure of the axoneme, revealing the tubulin content. The presence of hyaluronidase and tubulin is essential for the sperm function, and the analysis of the DNA status reveals the eventual apoptotic process. Using this method in normal spermatozoa, the staining of the acrosomal hyaluronidase reveals, by yellow-green fluorescence, the shape of the acrosomal complex and its texture. At the same time, in the same sperm cell, the staining of the axonemal tubulin demonstrates, by a red labeling, the presence of the protein and therefore the consistence of the axonemal structure. Simultaneously, at the head level, the absence of red labeling from nuclear DNA indicates that the apoptotic process is not present. This protocol allows quantification of the frequency of the presence of normal or abnormal spermatozoa, by an easy scoring and calculation of the apoptotic sperm or of the sperm with generic defects at acrosomal or flagellar level. The percentage of normal spermatozoa evaluated by the triple staining method has been compared with the results of the PAP staining and of the ultrastructural analysis, statistically elaborated. Triple staining results more severe than the PAP method, but TEM analysis is the finest technique to detect sperm abnormality because it considers the entire panel of sperm defects.
Human spermatozoa; Immunofluorescence; Triple staining; TUNE
10, 15 reciprocal translocation in an infertile man: ultrastructural and fluorescence in-situ hybridization sperm study: Case report
No full textBackground: Peculiar sperm defects are described in a sterile man heterozygous for a balanced translocation t(10;15) (q26;q12). As this structural reorganization was absent in the parents, the translocation must have appeared de novo in the present patient. Methods: Spermatozoa were analysed under light and transmission electron microscopy (TEM). Fluorescence in-situ hybridization (FISH) was performed on the lymphocyte karyotype. Aneuploidy frequencies of chromosomes 18, X and Y in sperm nuclei, not involved in the translocation, were investigated using three-colour FISH. Dual-colour FISH was used to evaluate segregation of chromosomes 10, 15 in decondensed sperm nuclei. Moreover, three-colour FISH, using telomeric probes for chromosomes 10, 15 was performed in order to distinguish balanced and unbalanced gametes. Results and conclusions: Overall, structural characteristics indicate general immaturity of the germinal cells. FISH sperm analysis detected an increase in chromosome 18 disomy (0.81%) suggesting an interchromosomal effect. A high frequency of diploidies, particularly 18,18,X,X and 18,18,X,Y, was also found. FISH segregation analysis for chromosomes 10, 15 indicated that 32.8% were balanced gametes, whereas 68.2% were unbalanced. Taken together, these data demonstrate in a male carrier of a reciprocal translocation t(10;15) the presence of diffuse ultrastructural sperm alterations and a high frequency of sperm aneuploidies. The existence of a correlation among these factors is proposed
Protein modification as oxidative stress marker in normal and pathological human seminal plasma
No full textObjective: Our study aims to assess the oxidative stress status of seminal plasma from normozoospermic, azoospermic, and leukocytospermic males, since abnormal sperm and leukocytes in human ejaculates are the main source of reactive oxygen species (ROS) which lead to oxidative damages. For this purpose we applied a biochemical approach to the assessment of the oxidative stress status by using twodimensional (2D) electrophoresis to check the level of protein oxidation after specific labeling of free thiol (-SH) groups. Methods: Seminal plasma samples from normal and pathological males were analyzed by a luminol-based chemiluminescent assay. The same samples after specific labeling of free -SH groups with 3-Nmaleimidopropionyl biocytin, were analyzed by 2D electrophoresis and computer-assisted semiquantitative determination of the amount of free -SH groups. Results: Using a standard chemiluminescence assay, we demonstrated a high, low and normal level of ROS, respectively, in seminal plasma from leukocytospermic, azoospermic, and normozoospermic subjects. By 2D electrophoresis and streptavidin blotting of specifically labeled free -SH groups of proteins, we detected in the same samples a higher level of oxidated -SH groups comparable between azoospermic and leukocytospermic samples, whereas a significantly higher level of free -SH groups was detected in normozoospermic subjects. Discussion: Our results demonstrated that a pathological oxidative stress status in seminal plasma may be revealed by the levels of the protein free -SH groups, both in the presence or absence of cells. © W.S. Maney & Son Ltd 2012
Localization of lamins in mammalian spermatozoa
No full textLittle information is available concerning lamins in the nucleus of germinal cells. In this paper we briefly describe and compare the organization of A- and B-type lamins in several mammalian spermatozoa. Nuclear lamin B is localized primarily in the postacrosomal sheath of human, bull and rabbit spermatozoa; lamin A/C is a major component of the equatorial segment in most of mammalian sperm, with the exception of rodents. In mouse sperm, devoid of equatorial segment, only lamin B appears to be expressed. The same happens in human pathological spermatozoa in which the equatorial segment is altered or absent
Different applications of the ultrastructural-mathematical evaluation of the sperm quality.
No full textIntracytoplasmic sperm injection and pregnancy with decapitated sperm
No full textObjective: To perform intracytoplasmic sperm injection (ICSI) in couples with primary infertility owing to sperm defects causing total immotility. Design: Case report. Setting: Couple Sterility Center, University of Siena. Patient(s): Two infertile couples, the male members of which had "detached tail" genetic sperm defect. Main Outcome Measure(s): Physical and hormonal assays, semen analysis by light and electron microscopy, Y microdeletion screening, immunofluorescence, fluorescence in situ hybridization analysis of sperm nuclei, and PCR for partial sequences of AKAP4/AKAP3 binding regions were performed. The couples then underwent ICSI. Result(s): Transmission electron microscopic analysis showed that the cause of sterility was "detached tail" genetic sperm defect. Immunofluorescence staining confirmed sperm structural alterations. Screening of Y microdeletions, partial sequences of AKAP4/AKAP3 binding regions, and fluorescence in situ hybridization did not show any sperm nucleus abnormalities. Three and two ICSI cycles were performed in the two couples. One pregnancy was achieved and a healthy baby with a normal female karyotype was born. Conclusion(s): One couple successfully underwent ICSI with "detached tail" sperm and gave birth to a healthy baby, suggesting that this structural abnormality may be bypassed by injecting sperm with a normal centriolar region. © 2010 American Society for Reproductive Medicine
Effect of follicle-stimulating hormone on sperm quality and pregnancy rate
No full textAim: To evaluate the possible links between ultrastructural sperm quality and the clinical pregnancy rate in infertile males treated with FSH before intracytoplasmic sperm injection (ICSI). Methods: Forty-four infertile males with idiopathic oligo-asthenozoospermia were randomly allocated to the treated (n=24) and non-treated (control, n=20) groups. Semen analysis was carried out by light and transmission electron microscopy (TEM) before and 12 weeks after FSH therapy. ICSI was performed in all couples. Results: TEM revealed a significant improvement in sperm quality after FSH treatment, particularly in men with their partners achieving clinical pregnancy. The pregnancy rate was 33% in the treated group and 20% in the control. Conclusion: Results highlight a positive role of FSH therapy in infertile males before ICSI, which was correlated with an increased pregnancy rate in treated couples. We believe that improved sperm ultrastructure after FSH therapy could positively influence the quality and early stage of embryo development, thereby increasing the probability of embryo implantation
Successful multiple pregnancy achieved after transfer of frozen embryos obtained via intracytoplasmic sperm injection with testicular sperm from an AZFc-deleted man
No full textObjective: To describe a case of successful triplet pregnancy after testicular sperm extraction (TESE) from a man with AZFc deletion and intracytoplasmic sperm injection (ICSI). Design: Case report. Setting: University hospital. Patient(s): A 38-year-old man affected by complete AZFc deletion and azoospermia. Intervention(s): Spermiogram, Y-chromosome microdeletion screening, TESE for sperm recovery from testicular tissue on the same day as ICSI, transfer of frozen-thawed embryos, vaginal ultrasound examination. Main Outcome Measure(s): The Y chromosome genetic status of an azoospermic patient who underwent TESE and ICSI, the fertilization and pregnancy outcome. Result(s): The patient was found to be azoospermic, and the deletion screening showed complete AZFc deletion. After TESE, the recovered testicular sperm were selected for ICSI. Three good quality embryos were obtained and were frozen due to ovarian hyperstimulation syndrome in the female partner. After transfer of the thawed embryos, a triplet pregnancy was diagnosed by vaginal ultrasonography at the seventh week of gestation. Two male and one female healthy babies were born. Conclusion(s): This is the first report of a successful triplet pregnancy after the transfer of frozen-thawed embryos in a couple in whom the male partner was azoospermic and a carrier of complete AZFc deletion. This deletion should not adversely affect a man's TESE retrieval prognosis or the fertilization, cleavage, and implantation of embryos. The offspring were healthy, although the two sons inherited the AZFc deletion. Copyright © 2010 American Society for Reproductive Medicine, Published by Elsevier Inc
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