Navegando por Palavras-chave "spermatozoa"
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- ItemSomente MetadadadosEffect of varicocele on sperm function and semen oxidative stress(Wiley-Blackwell, 2012-01-01) Blumer, Camile Garcia [UNIFESP]; Restelli, Adriana Ester [UNIFESP]; Del Giudice, Paula Toni [UNIFESP]; Soler, Thiesa Butterby [UNIFESP]; Fraietta, Renato [UNIFESP]; Nichi, Marcilio; Bertolla, Ricardo Pimenta [UNIFESP]; Cedenho, Agnaldo Pereira [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP)OBJECTIVETo assess the effect of varicocele on sperm DNA integrity, mitochondrial activity, lipid peroxidation and acrosome integrity.PATIENTS and METHODSIn all, 30 patients with a clinically diagnosed varicocele of grade II or III and 32 men without a varicocele were evaluated for sperm DNA fragmentation (comet assay), mitochondrial activity (3,3'-diaminobenzidine assay), lipid peroxidation (malondialdehyde) and acrosome integrity (fluorescent probe labelled peanut agglutinin).RESULTSThe varicocele group showed fewer spermatozoa with intact DNA (grade II, P = 0.040), more cells with inactive mitochondria (class III, P = 0.001), fewer cells with active mitochondria (class I, P = 0.005) and fewer spermatozoa with intact acrosomes (P < 0.001). Finally, no significant differences were observed in lipid peroxidation levels.CONCLUSIONMen with varicocele showed an increase in sperm DNA fragmentation and a reduction in mitochondrial activity and acrosome integrity. However, lipid peroxidation levels remained unchanged.
- ItemAcesso aberto (Open Access)Effects of the technique of cryopreservation and dilution/centrifugation after thawing on the motility and vitality of spermatozoa of oligoasthenozoospermic men(Sociedade Brasileira de Urologia, 2003-04-01) Esteves, Sandro C. [UNIFESP]; Spaine, Deborah M. [UNIFESP]; Cedenho, Agnaldo Pereira [UNIFESP]; Srougi, Miguel [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)OBJECTIVE: Comparing in human semen samples with low initial quality, the effects of 2 techniques of cryopreservation and dilution/centrifugation after thawing on the spermatic motility and vitality. MATERIALS AND METHODS: Semen samples from 15 oligo and/or asthenozoospermic individuals assisted in the infertility sector of a tertiary hospital were obtained through masturbation. The samples were divided into 2 portions of equal volume, and diluted (1:1; v/v) with the cryoprotector containing glycerol (Test yolk buffer). One portion was frozen through the technique of liquid nitrogen vapor with static phases (group I - GI), while the other was frozen through a programmable biological freezer with linear speed (Planer, Kryo 10, series III) (group II - GII). The following parameters were assessed before freezing and after thawing: percentage of spermatozoa with progressive motility (Prog%) and percentage of live spermatozoa (Vit%). After defrosting, Prog% was assessed before and after removal of cryoprotector diluent, in different time intervals (zero, 3 h, and 24 h). The statistical analysis has been accomplished by using the non-parametric tests of Wilcoxon and Friedman. RESULTS: There was significant reduction of Prog% and Vit% from before freezing to after defrosting in both groups, I and II (p < 0.001). Values of Prog% and Vit% were not statistically different between groups, after thawing. It has been observed a significant reduction in Prog% among portions frozen with the automated technique after dilution and centrifugation for removal of cryoprotector (p = 0.006). After cryoprotector removal, Prog% has been kept unaltered, in both groups, during the first 3 hours of incubation, although being superior in group I (p = 0,04). There was a significant decrease in Prog% after 24 hours of incubation, in both groups (p < 0,01). CONCLUSION: For human semen samples with low initial quality, freezing through vapor technique or through the automated technique showed to be equivalent in regarding recovery of live spermatozoa with progressive motility. The effects of dilution and centrifugation to remove the cryoprotector had a negative impact only in samples frozen through the automated technique. In both techniques, progressive motility is kept constant during the first 3 hours after thawing and removal of the cryoprotector, but is drastically diminished by the end of an incubation period of 24 hours.
- ItemSomente MetadadadosThe efficacy of repeat percutaneous epididymal sperm aspiration procedures(Lippincott Williams & Wilkins, 2003-05-01) Pasqualotto, F. F.; Rossi-Ferragut, L. M.; Rocha, C. C.; Iaconelli, A.; Ortiz, V; Borges, E.; Fertil Ctr Assisted Fertilizat; Universidade Federal de São Paulo (UNIFESP)Purpose: the development of intracytoplasmic sperm injection spawned new methods of sperm retrieval for men with obstructive azoospermia who did not want to undergo reconstruction of the seminal tract. There is a wide array of different procedures that may be performed in these cases, for instance percutaneous epididymal sperm aspiration (PESA) and microepididymal sperm aspiration. However, concerns regarding the presence of sperm in a second PESA attempt due to possible fibrosis have been suggested by many authors. We evaluate if it is worthwhile to repeat percutaneous epididymal sperm aspiration.Materials and Methods: the records of 20 patients (23 attempts) who underwent repeat PESA from January 1996 to September 2000 for assisted reproductive technique purposes were reviewed. in all patients the repeat procedure was performed on the same side as the previous PESA. Data were collected on patient age, presence of motile sperm during PESA, epididymal side, pregnancies and abortion rates.Results: Mean patient age +/- SD was 32.4 +/- 5.6 years. One patient was excluded from our analysis due to lack of information on the chart regarding the side of the procedure. Repeat PESA was performed in the right epididymis in 12 attempts and in the left in 10. of the remaining 19 patients 14 (73.68%) did not and 5 (26.3%) have sperm in the epididymal fluid. in these 5 patients 8 repeat PESA procedures were performed (3 procedures in 1 and motile sperm was always found (8 of 22 attempts, 36.4%). Three patients achieved pregnancy with the motile sperm retrieved from the repeat PESA (3 of 8 repeat attempts, 37.5%). No abortions were detected.Conclusions: More than a third of repeat PESA attempts resulted in the presence of motile sperm. Before performing testicular sperm aspiration or extraction in patients who have undergone previous PESA without achieving pregnancy, repeat PESA may be done. Further attempts should be added in the future to confirm these results.
- ItemSomente MetadadadosEtiology-specific outcomes of intracytoplasmic sperm injection in azoospermic patients(Elsevier B.V., 2005-03-01) Pasqualotto, Fábio F.; Rossi, Lia Mara; Guilherme, Patricia; Ortiz, Valdemar [UNIFESP]; Iaconelli, Assumpto; Borges Junior, Edson; Fertil Ctr Assisted Reprod; Jundiai Med Sch; Universidade Federal de São Paulo (UNIFESP)Objective: To assess fertilization, pregnancy, and miscarriage rates after intracytoplasmic sperm injection (ICSI) with epididymal or testicular spermatozoa from different types of azoospermia.Design: Retrospective study.Setting: Academic medical center and private fertility center.Patient(s): Two hundred twelve patients underwent 257 ICSI cycles.Intervention(s): Cycles of ICSI were divided into four groups according to the etiology of azoospermia: A (nonobstruction), B (postvasectomy), C (congenital obstruction), and D (obstruction due to infection). Testicular sperm aspiration and percutaneous epididymal sperm aspiration were the sperm retrieval methods used for ICSI.Main Outcome Measure(s): Fertilization, pregnancy, and miscarriage rates.Result(s): Normal fertilization rates were higher in groups C (67.7%) and B (64.1%) compared with groups A (47.3%) and D (58.9%). Although lower pregnancy rates were seen in group A, no statistical differences were detected among groups. However, the miscarriage rate was higher in group A (45.6%) compared with groups B (25.25%), C (24%), and D (22.58%).Conclusion(s): Although no differences were detected in the pregnancy rates across groups, fertilization and implantation rates were higher in patients with congenital obstruction of the seminal path. the pregnancy rate was higher and the miscarriage rate lower when epididymal sperm was used compared with testicular sperm. (c) 2005 by American Society for Reproductive Medicine.
- ItemAcesso aberto (Open Access)Late reproductive analysis in rat male offspring exposed to nicotine during pregnancy and lactation(Wiley-Blackwell, 2016) Miranda-Spooner, M. [UNIFESP]; Paccola, C. C. [UNIFESP]; Neves, F. M. O. [UNIFESP]; de Oliva, S. U. [UNIFESP]; Miraglia, S. M. [UNIFESP]We previously observed that nicotine, administered to rats (Wistar) during pregnancy and lactation periods, provokes, in the progeny, late morphofunctional alterations in Leydig cell, body weight increase in adulthood (90days post partum, dpp) as well as seminiferous epithelium injury. Aiming to investigate whether the spermatogenic damage previously observed in adult progenies from pregnant and lactating nicotine-exposed rat dams are maintained or whether it is worsened in older rats, we analyzed the morphological testicular alterations after up to two complete periods of spermatogenesis (53days each), spermatic parameters, and sperm DNA fragmentation. Pregnant and lactating rats were nicotine-exposed (2mg/kg/day) through an osmotic minipump implanted on the first day of pregnancy and replaced after birth. Absolute Control (no minipump) and Sham Control (minipump without nicotine) groups were established. The offspring were killed at 90, 143, and 196dpp. Significant alterations in morphometric and stereological testicular parameters, such as concentration of sperm number, daily sperm production, and plasma and intratesticular levels of cholesterol and testosterone were not observed in nicotine-exposed rats. Testicular histopathological analysis showed small intraepithelial vacuolization and an accentuated germ cell desquamation in exposed rats. However, the offspring from nicotine-exposed dams exhibited higher frequency of morphologically abnormal spermatozoa and lower sperm motility in comparison with control groups. In addition, nicotine-exposed groups showed a significant reduction in sperm mitochondrial activity and an increased sperm DNA fragmentation (Comet assay). These results indicate a late reproductive damage in the male progeny caused by maternal nicotine exposure, related to the decrease in sperm quality.
- ItemAcesso aberto (Open Access)Search and identification of spermatozoa and spermatids in the ejaculate of non-obstructive azoospermic patients(Sociedade Brasileira de Urologia, 2005-02-01) Timm Jr, Odival [UNIFESP]; Cedenho, Agnaldo Pereira [UNIFESP]; Spaine, Deborah M. [UNIFESP]; Buttignol, Marcia H.p. [UNIFESP]; Fraietta, Renato [UNIFESP]; Ortiz, Valdemar [UNIFESP]; Srougi, Miguel [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)OBJECTIVE: To search and to identify spermatozoa and spermatids, present in the ejaculate of non-obstructive azoospermic patients. MATERIALS AND METHODS: 27 patients, aged between 18 and 48 years, with initial diagnosis compatible with non-obstructive azoospermia, underwent up to 3 seminal samples, with assessment of macroscopic and microscopic parameters differentiated for each sample. In the first sample, 5 µL of semen were analyzed in a Horwell chamber in order to assess the presence or absence of spermatozoa. The procedure was repeated with 2 other aliquots. In the absence of spermatozoa, the entire sample was transferred to a conic tube and following centrifugation the sediment was freshly analyzed. The second seminal sample was collected only when no spermatozoa were found in the first sample and the research was performed in the same way. In cases where spermatozoa were not seen, the sample was centrifuged and the obtained sediment was stained by the panoptic method and observed under common light microscopy (1250X). The third seminal sample was collected only in cases when patients had not shown spermatozoa in the first and second seminal samples. RESULTS: 4/27 (14.8%) patients presented spermatozoa in the first seminal sample and 6/23 (26.1%), in the second seminal sample. No spermatozoa were seen in the third sample, however, 11/17 (64.7%) presented spermatids. CONCLUSION: In clinical situations where the initial diagnosis is non-obstructive azoospermia, one single routine seminal analysis is not enough to confirm this diagnosis and the analysis of the centrifuged sediment can have relevant clinical consequences. Among patients considered non-obstructive azoospermic, when duly assessed, 37% presented spermatozoa and 64.7%, spermatids.
- ItemSomente MetadadadosSerotonin concentration, synthesis, cell origin, and targets in the rat caput epididymis during sexual maturation and variations associated with adult mating status: Morphological and biochemical studies(Amer Soc Andrology, Inc, 2007-01-01) Jimenez-Trejo, Francisco; Tapia-Rodriguez, Miguel; Queiroz, Daniel Barboza Cava [UNIFESP]; Padilla, Patricia; Avellar, Maria Christina Werneck [UNIFESP]; Manzano, Patricia Rivas; Manjarrez-Gutierrez, Gabriel; Gutierrez-Ospina, Gabriel; Natl Autonomous Univ Mexico; Universidade Federal de São Paulo (UNIFESP); XXI Century Natl Med CtrThe caput epididymis of some mammals contains large quantities of serotonin whose origin, targets, and physiological variations have been poorly studied. We combined morphological and biochemical techniques to begin approaching these aspects of serotonin in the rat caput epididymis. Serotonin immunostaining was detected in mast, epithelial, and neuroendocrine cells. Epithelial cells displayed immunoreactivity to 5HT(1A), 5HT(2A), and 5HT(3) serotonin receptors. Endothelial and mast cells labeled positive for 5HT(1B) serotonin receptors and spermatozoa displayed 5HT2A and 5HT3 serotonin receptor immunoreactivity. Epithelial, endothelial, and mast cells stained positive for serotonin transporters. Only epithelial cells showed tryptophan hydroxylase immunoreactivity; this enzyme catalyzes the limiting step in the serotonin synthetic pathway. in addition, Western blot analyses of caput homogenates documented the presence of 2 protein bands (approximate to 51 kd and 48 kd) that were immunoreactive for tryptophan hydroxylase. Chromatographic analyses documented the presence of tryptophan hydroxylase in the caput, and showed that both its activity and serotonin availability increased with sexual maturation and decreased following p-chlorophenylalanine treatment, an inhibitor of tryptophan hydroxylase activity. Interestingly, serotonin concentration and tryptophan hydroxylase activity tended to be higher in breeding males than in those with no mating experience. We think that these results support the existence of a local serotoninergic system in the rat caput epididymis that might regulate some aspects of male reproductive function.
- ItemAcesso aberto (Open Access)Single nucleotide polymorphisms of the heat shock protein 90 gene in varicocele-associated infertility(Sociedade Brasileira de Urologia, 2005-06-01) Hassun Filho, Pericles A. [UNIFESP]; Cedenho, Agnaldo Pereira [UNIFESP]; Lima, Samira B. [UNIFESP]; Ortiz, Valdemar [UNIFESP]; Srougi, Miguel [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)PURPOSE: Varicoceles are associated with impaired testicular function and male infertility, but the molecular mechanisms by which fertility is affected have not been satisfactorily explained. Spermatogenesis might be affected by increased scrotal temperature, such as that caused by varicocele. HSP90 is a molecular chaperone expressed in germ cells and is related to spermatogenesis, motility, and both heat and oxidative stress. Possible correlations between coding single region nucleotide polymorphisms (cSNPs) in the HSP90 gene in patients with varicocele associated with infertility were analyzed, and polymorphisms in these exons were characterized through DNA sequencing. MATERIALS AND METHODS: PCR-SSCP and DNA sequencing were used to search for mutations in 18 infertile patients with varicocele, 11 patients with idiopathic infertility and 12 fertile men. DNA was extracted from leucocytes for PCR amplification and SSCP analysis. DNA from samples with an altered band pattern in the SSCP was then sequenced to search for polymorphisms. RESULTS: Three silent polymorphisms that do not lead to amino acid substitutions were identified. CONCLUSION: Mutations in the HSP90 gene do not appear to be a common cause of male factor infertility. The low incidence of gene variation, or SNPs, in infertile men demonstrates that this gene is highly conserved and thus confirms its key role in spermatogenesis and response to heat stress.