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- ItemSomente MetadadadosAnálise do polimorfismo de nucleotídeo único (SNP)RS2071307 selecionado no gene da elastina (ELN) em indivíduos do gênero masculino portadores de hérnia inguinal do tipo IIIa da Classificação de Nyhus(Universidade Federal de São Paulo (UNIFESP), 2020-11-26) Oliveira, Guilherme Blattner Torres De [UNIFESP]; Lopes Filho, Gaspar De Jesus [UNIFESP]; Universidade Federal de São PauloObjectives: To evaluate the frequency of the single nucleotide polymorphism (SNP) rs2071307 in the ELN gene in male individuals with type IIIa inguinal hernia from the Nyhus classification, as well as to verify the association of this polymorphism with a positive family history in first-degree relatives degree for inguinal hernia. Methods: This is a case-control, multicenter and multidisciplinary study with 47 individuals submitted to inguinal herniorrhaphy and 50 individuals of the same age group, without a diagnosis of inguinal hernia. For this study, peripheral venous blood was collected from each individual, DNA extraction and genotyping by real-time PCR. Descriptive statistics included calculations of mean, median and standard deviation, as well as absolute and relative frequencies for categorical variables. Chi-square and Fisher's exact tests were used to assess the association between categorical variables. The Hardy-Weinberg balance was tested for the SNP in the case and control groups, as well as the chi-square test. To evaluate the association of the polymorphism and the occurrence of inguinal hernia, Odds ratios and the respective 95% confidence intervals were estimated. For all statistical tests, the results were considered statistically significant when p <0.05. All statistical analyzes were performed using the software Stata version 15.1. Results: The mean and median age of individuals with inguinal hernia were 48.3 and 40 years respectively (standard deviation: 14.1 years), whereas in controls, they were 40.4 and 50 years (standard deviation: 11, Four years). The genotypes of the groups were found in Hardy- Weinberg Equilibrium. The rs2071307 polymorphism and its respective GG, GA and AA genotypes showed a similar distribution between cases and controls (p = 0.499), with a predominance of the heterozygous GA genotype. The G allele was more frequent, being present in 86% of the sample between cases and controls. There was no statistical association between family history in first-degree relatives and inguinal hernia, both in cases with controls (p = 0.502 and p = 0.861, respectively). Conclusions: The rs2071307 polymorphism of the ELN gene, with its respective genotypes, was not associated with type IIIa inguinal hernia of the Nyhus classification, just as there was no association between this polymorphism and the presence of positive family history in first degree relatives for hernia. inguinal.
- ItemSomente MetadadadosA comparative study of extracellular matrix remodeling in two murine models of emphysema(F Hernandez, 2013-02-01) Lopes, F. D. T. Q. S.; Toledo, A. C.; Olivo, C. R.; Prado, Carla Maximo [UNIFESP]; Leick-Maldonado, Edna Aparecida [UNIFESP]; Medeiros, M. C.; Santos, A. B. G.; Garippo, A.; Martins, M. A.; Mauad, T.; Universidade de São Paulo (USP); Universidade Federal de São Paulo (UNIFESP)A single instillation of porcine pancreatic elastase (PPE) results in significant airspace enlargement on the 28th day after instillation, whereas cigarette smoke (CS) exposure requires 6 months to produce mild emphysema in rodents. Considering that there are differences in the pathogenesis of parenchymal destruction in these different experimental models, it is likely that there may be different patterns of extracellular matrix (ECM) remodeling. To evaluate ECM remodeling, C57BL/6 mice were submitted to either a nasal drop of PPE (PPE 28 Days) or exposed for 6 months to cigarette smoke (CS 6 months). Control groups received either an intranasal instillation of saline solution (Saline 28 Days) or remained without any smoke inhalation for six months (Control 6 months). We measured the mean linear intercept and the volume proportion of collagen type I, collagen type III, elastin and fibrillin. We used emission-scanning confocal microscopy to verify the fiber distribution. Both models induced increased mean linear intercept in relation to the respective controls, being larger in the elastase model in relation to the CS model. In the CS model, emphysema was associated with an increase in the volume proportion of fibrillin, whereas in the PPE model there was an increase in the parenchymal elastin content. In both models, there was an increase in collagen type III, which was higher in the CS-exposed mice. We concluded that ECM remodeling is different in the two most used experimental models of emphysema.
- ItemAcesso aberto (Open Access)Expressão de genes e de proteínas envolvidos na biossíntese da matriz extracelular no tecido vaginal de mulheres com e sem prolapso de órgãos pélvicos(Universidade Federal de São Paulo (UNIFESP), 2011-05-25) Bortolini, Maria Augusta Tezelli [UNIFESP]; Sartori, Marair Gracio Ferreira [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Objective: Pelvic organ prolapse (POP) results from the failure of pelvic floor support, and connective tissue abnormalities may be involved in the etiology and/or progression of the dysfunction. We aimed to analyze the differential expression of genes and proteins related to the collagen and elastin biogenesis: lysyl oxidases (LOXs), fibulin-5, fibrillin -1 and -2, and procollagen C proteinase (PCP/BMP1) in vaginal tissue of women without and with advanced POP controlled by hormonal status. Materials and Methods: During total hysterectomy, anterior vaginal wall biopsies were obtained from Caucasian premenopausal women (proliferative phase of menstrual cycle) and postmenopausal women with severe POP (POPQ stage III and IV) and asymptomatic controls (POPQ 0). Total mRNA and protein were extracted using Trizol and RIPA buffer, and the genes and proteins of interest were quantified by real-time RT-PCR and Immunoblotting, respectively. The following analysis were performed: (1) expression of LOX family genes and proteins (LOX and LOXL1-4), fibulin-5, fibrillin-1 and -2 in premenopausal women with and without POP; (2) PCP/BMP1 gene and protein expression in vaginal tissue of pre- and postmenopausal POP women, and respective controls. Wilcoxon signed-rank and Fisher tests were used for statistical analysis (p<0.05). Results: Samples from 15 premenopausal patients and 11 controls were obtained for study (1); 39 premenopausal (POP=23 and Control=16) and 18 postmenopausal women samples (POP=13 and Control=5) for study (2). We observed: (1) significant decrease in expression of LOX, LOXL1 and LOXL3 genes, as well as LOX and LOXL3 proteins in vaginal tissue of premenopausal POP patients compared with asymptomatic women (p<0.05); (2) PCP/BMP1 gene downregulation in the vagina of women with severe POP compared with controls, in both premenopausal and postmenopausal phase (both p=0.01); significant reduction of 130 kDa, 92.5 kDa and 82.5 kDa PCP/BMP1 isoforms in vaginal tissue of postmenopausal patients (p=0.01), and 130 kDa isoform upregulation in premenopausal women with severe POP (p=0.009), compared with their respective controls. Conclusion: The expression of LOXs enzymes and PCP/BMP1 are altered in vaginal tissue of women with severe POP, and are modulated by hormonal status. Dysregulation of these enzymes involved in the extracellular matrix biogenesis may contribute to impaired tissue and vaginal support, and may be involved in POP development.
- ItemSomente MetadadadosGenetics of pelvic organ prolapse: crossing the bridge between bench and bedside in urogynecologic research(Springer, 2011-10-01) Bortolini, Maria Augusta Tezelli [UNIFESP]; Rizk, Diaa E. E.; Universidade Federal de São Paulo (UNIFESP); Ain Shams UnivAn increasing number of scientists have studied the molecular and biochemical basis of pelvic organ prolapse (POP). the extracellular matrix content of the pelvic floor is the major focus of those investigations and pointed for potential molecular markers of the dysfunction. the identification of women predisposed to develop POP would help in the patients' management and care. This article includes a critical analysis of the literature up to now; discusses implications for future research and the role of the genetics in POP.
- ItemAcesso aberto (Open Access)Imunohistoquímica como método de estudo das fibras elásticas em prega vocal humana(ABORL-CCF Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial, 2005-08-01) Ramos, Hugo Valter Lisboa [UNIFESP]; Simões, Manuel de Jesus [UNIFESP]; Pontes, Paulo Augusto de Lima [UNIFESP]; Neves, Luciano Rodrigues [UNIFESP]; Barbosa, Luiz Henrique Fonceca; De Biase, Noemi Grigoletto [UNIFESP]; Oshima, Celina Tizuko Fujiyama [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)AIM: Verify the use of immunohistochemistry as a method to measure all forms of elastic fibers at human vocal folds. STUDY DESIGN: transversal cohort. MATERIAL AND METHOD: We collected vocal folds following these criterion: age between 25 and 40, Caucasian men, dead by gun shot, within 12 hours of death, without instrumentation of the larynx or suspicion of neck injury and without mucosal lesions noted by microscopy. Ten vocal folds were collected and one, of a man aged 28 years, was selected to study. The vocal fold was transversely cut in 9 regions and in each segment three slides were made. These slides were stained by Verhoeff and Weighert's resorcin-fuchsin and used for immunohistochemistry. The elastic compound was measured by colorimetric software analysis. RESULTS: In Verhoeff and Weighert's resorcin-fuchsin, the intermediate and deep layer showed values higher than those of the superficial layer. The amount of tropoelastin identified by the antibody at the superficial layer was close to those of intermediate and deep layer. CONCLUSION: Immunohistochemistry is a method that can identify and measure all forms of elastic fibers at human vocal fold.