Navegando por Palavras-chave "acrosome"

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    Análise dos aspectos funcionais dos espermatozoides e do perfil proteômico de plasma seminal de homens tabagistas
    (Universidade Federal de São Paulo (UNIFESP), 2014-02-12) Antoniassi, Mariana Pereira [UNIFESP]; Bertolla, Ricardo Pimenta Bertolla [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Objectives: To evaluate the effect of smoking on sperm functional aspects, semen oxidative stress and seminal plasma proteomic profile. Method: A cross-sectional observational study including 20 nonsmokers patients with normal semen quality, according to the World Health Organization (2010) (control group), and 20 smoking patients (study group). Beyond semen analysis, sperm function tests were performed on each patient: evaluation of DNA integrity by alkaline Comet assay; analysis of mitochondrial activity using 3,3' - diaminobenzidine (DAB) staining; and evaluation of acrosomal integrity by PNA (Peannut Agglutinin) binding to the outer acrosomal membrane. The seminal plasma was used to assess the level of oxidative stress by TBARS (Thiobarbituric acid reactive substances) quantification and label-free quantitative proteomic analysis by LC-MS/MS. Groups were compared using a Student?s T test (p <0.05), considering a minimum fold-change of 2.0 (or maximum of 0.5). Results: The group of smokers presented higher percentage of DNA damage (Comet class III and IV), partially and fully inactive mitochondria (DAB class III and IV) and non-intact acrosomes when compared to the control group. No difference in the level of oxidative stress among groups was observed. With respect to proteomic analysis, 422 proteins were identified and quantified, of which one protein was absent (Mammaglobin-B), 6 proteins were underexpressed (Prolargin, Poly(rC)- binding protein 1, Bile salt-activated lipase, Carbonyl reductase [NADPH] 1, Phospholipase A2, membrane associated, CD44 antigen) and 3 proteins were overexpressed among smokers (S100-A9, Neutrophil gelatinase-associated lipocalin e Alpha-1-acid glycoprotein 1). Conclusions: Cigarette smoking alters the functional quality of sperm and the seminal plasma proteomic profile of men, but does not alter the levels of oxidative stress. The overexpressed proteins have functions related to the metabolism of steroids and derivatives of arachidonic acid, as well as preservation of the extracellular scaffold. Among the overexpressed proteins, we highlight the immune function.
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    Analysis of the functional aspects and seminal plasma proteomic profile of sperm from smokers
    (Wiley, 2016) Antoniassi, Mariana Pereira [UNIFESP]; Intasqui, Paula [UNIFESP]; Camargo, Mariana [UNIFESP]; Zylbersztejn, Daniel Suslik [UNIFESP]; Carvalho, Valdemir Melechco; Cardozo, Karina H. M.; Bertolla, Ricardo Pimenta [UNIFESP]
    Objective To evaluate the effect of smoking on sperm functional quality and seminal plasma proteomic profile. Patients and Methods Sperm functional tests were performed in 20 non-smoking men with normal semen quality, according to the World Health Organization (2010) and in 20 smoking patients. These included: evaluation of DNA fragmentation by alkaline Comet assay
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    Effect 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.
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    O impacto da obesidade sobre a qualidade funcional dos espermatozoides e os mecanismos proteômicos do sêmen
    (Universidade Federal de São Paulo (UNIFESP), 2015-11-30) Ferigolo, Paola Cristine [UNIFESP]; Fraietta, Renato Fraietta [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Purpose: To evaluate the effect of obesity on the sperm function and seminal plasma proteome. Methods: A prospective study was performed including 47 male volunteers, of which 27 participated in the obese group (BMI ? 30 kg / m2) and 20 in the control group (? 18.5 and BMI <25 kg / m2). After semen retrieval by masturbation, an aliquot was used for semen analysis and another for evaluation of the main functional characteristics of sperm. Mitochondrial activity was determined by a colorimetric stain (3,3?-diaminobenzidine - DAB) and classified as DAB I (all mitochondria active) to DAB IV (all mitochondria inactive); acrosome integrity was determined by a fluorescent probe (PNA-FITC) and assessed by fluorescent microscopy; and sperm DNA fragmentation was assessed by an alkaline Comet assay, in which sperm were classified as high DNA integrity (class I) to high DNA fragmentation (class IV). The remaining semen volume was centrifuged and the supernatant seminal plasma was utilized for proteomic analysis by LC-MS/MS. Groups were compared by an unpaired Student?s t test. Differentially expressed proteins were subsequently used for functional enrichment analysis of Gene Ontology categories using the online platform PantherDB. Results: Significant diferences were observed in sperm non-progressive motility (p = 0.018), morphology (p = 0.028), PNA-FITC (p <0.0001), DAB I (p = 0.0001), DAB III (p = 0 , 0017), IV DAB (0.026), DAB (p = 0.0003), Comet III (p = 0.001), and Comet (p = 0.003). In proteomics analysis, a total of 489 proteins were quantified and identified, in which 73 are differentially expressed between the two groups and 416 proteins are conserved. Among the differentially expressed proteins, 1 protein was absent, 19 were down-regulated, 49 were up-regulated and 4 were exclusive in the study group. The main functions enriched in the obesity group were negative regulation of the intrinsic pathway of apoptosis, regulation of phagocytosis, endocytosis regulation, regulation of proteolysis, endopeptidase inhibitory activity, activation of immune and inflammatory and antioxidant activity. The functions depleted in the obesity group were the following: regulation of metabolic process, regulation of response to stimulus and regulation of cellular process. Conclusions: Based on our findings, obesity may affect semen quality and sperm function. There is a significant change in the proteome of the seminal plasma of men with excess weight that elucidates some processes involved in infertility of these men, such as excessive production of reactive oxygen species, endocrine disorders, testicular hyperthermia, exacerbated immune response, inflammatory response, inhibiting phospholipase and changes in apoptotic pathway.