FERNANDA GUEDES LUIZ
Projetos de Pesquisa
Unidades Organizacionais
LIM/56 - Laboratório de Investigação em Dermatologia e Imunodeficiências, Hospital das Clínicas, Faculdade de Medicina
5 resultados
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conferenceObject Confocal Microscopy Image Analysis of Pancreatic beta-Cells K-ATP Channel Proteins in Congenital Hyperinsulinism (CHI)(2012) LOVISOLO, S. M.; GARIPPO, A. L.; GUEDES, F.; ZERBINI, M. C.Background: CHI is a life-threatening disorder of glucose metabolism of neonates characterized by serum insulin levels unresponsive to blood glucose concentrations. Pancreatic ß-cells regulates insulin secretion through ATP sensitive potassium channels (KATP channel) formed by sulfonylurea receptor 1 (SUR1) and potassium inward rectifying 6.2 (Kir 6.2) proteins. The ß-cell K ATP channel dependent CHI is classically associated with loss-of-function mutations of these subunits genes. In a previous study [1], no mutations in the Kir6.2 gene and in the 33-37 exons hot spot region of the SUR1 gene were identified in these patients. The aim of this study is to investigate if these subunits were present in the ß-cells of CHI patients. Design: Pancreatic surgical paraffin tissue from 7 neonates (3F/4M, 4-13 mo) with CHI and 8 autopsy pancreatic control tissue (5F/3M, 4-11 mo) were included in the study. Confocal microscopy double-staining immunofluorescence (insulin/SUR1 and insulin/Kir6.2) was performed in order to detect each protein specifically in the ß-cells All sections were analyzed under a Zeiss 510 Meta confocal laser scanning microscope (63x). At least 10 different endocrine islets were captured to evaluate the expression of either Kir6.2 or SUR1 (green-488nm) and insulin (red-633nm). Co-expression of insulin/SUR1 and insulin/Kir6.2 was analysed visually (green x red→yellow) and through correlation Pearson’s coefficient(PC) that estimates the goodness of rate association of the two fluorochromes. PC was compared among cases and controls using a nonparametric method (Mann-Whitney). Results: Visual observation clearly revealed the presence of Kir6.2 and SUR1 in a granular cytoplasmic pattern in the ß-cells of cases and controls. Nevertheless, overlap of insulin/Kir6.2 and insulin/SUR1 seemed to be more constant and uniform in controls than in CHI cases, confirmed by the analysis through PC showing a statistically significant decrease in Kir6.2 (P= 0.0084) and SUR1 (P= 0.041) in the ß-cells of CHI patients. Conclusions: This is the first demonstration of K ATP channels subunits Kir6.2 and SUR1 in the pancreatic ß-cells of CHI patients using an in situ method. Our results show that both subunits were present in the ß-cells, but are under-expressed in CHI patients compared to controls, adding a new information to this rare condition with a complex pathogenesis.- Immunohistochemical Evaluation of Skin Before and After Micro-Ablative Fractional Erbium-Doped Yttrium Aluminum Garnet Laser Treatment(2011) ODO, Lilian M.; ODO, Marina E. Y.; GUEDES, Fernanda; SOTTO, Mirian N.; CUCE, Luiz Carlos
- Inate immunity in rosacea. Langerhans cells, plasmacytoid dentritic cells, Toll-like receptors and inducible oxide nitric synthase (iNOS) expression in skin specimens: case-control study(2018) MOURA, Ana Karina Alves; GUEDES, Fernanda; RIVITTI-MACHADO, Maria Cecilia; SOTTO, Mirian N.Rosacea is a chronic inflammatory condition with predominant facial involvement. Because of that, many patients sense that rosacea affects quality of life. The etiology of rosacea remains unknown. Recent studies have suggested that aberrant innate immunity is central to this disease. The aim of this study was to examine the presence of Langerhans cells, plasmacytoid dentritic cells (PDC), the expression of Toll-like receptors (TLR) and inducible oxide nitric synthase (iNOS) in skin of patients with rosacea, to highlight the participation of innate immunity in its pathogenesis. 28 biopsy specimens were taken from patients with clinical and histopathological findings of rosacea. Immunohistochemical demonstration of Langerhans cells (anti-CD1a antibody), PDC (anti-CD 123 antibody), TLR2, TLR4 and iNOS was performed in skin samples and compared with normal skin controls. The expression of Langerhans cells was lower in rosacea group than in control group. PDC were found in skin samples of rosacea as isolated cells and forming small clusters. Expression of TLR2, TLR4 and iNOS was higher in rosacea samples than in normal skin controls. This research demonstrates early and late stage components of innate immunity in specimens of rosacea ratifying the existence of an altered innate immunity in its pathogenesis.
conferenceObject Confocal Microscopy Image Analysis of Pancreatic beta-Cells K-ATP Channel Proteins in Congenital Hyperinsulinism (CHI)(2012) LOVISOLO, S. M.; GARIPPO, A. L.; GUEDES, F.; ZERBINI, M. C.Background: CHI is a life-threatening disorder of glucose metabolism of neonates characterized by serum insulin levels unresponsive to blood glucose concentrations. Pancreatic ß-cells regulates insulin secretion through ATP sensitive potassium channels (K ATP channel) formed by sulfonylurea receptor 1 (SUR1) and potassium inward rectifying 6.2 (Kir 6.2) proteins. The ß-cell K ATP channel dependent CHI is classically associated with loss-of-function mutations of these subunits genes. In a previous study [1], no mutations in the Kir6.2 gene and in the 33-37 exons hot spot region of the SUR1 gene were identified in these patients. The aim of this study is to investigate if these subunits were present in the ß-cells of CHI patients. Design: Pancreatic surgical paraffin tissue from 7 neonates (3F/4M, 4-13 mo) with CHI and 8 autopsy pancreatic control tissue (5F/3M, 4-11 mo) were included in the study. Confocal microscopy double-staining immuno fluorescence (insulin/SUR1 and insulin/Kir6.2) was performed in order to detect each protein specifically in the ß-cells. All sections were analyzed under a Zeiss 510 Meta confocal laser scanning microscope (63x). At least 10 different endocrine islets were captured to evaluate the expression of either Kir6.2 or SUR1 (green-488nm) and insulin (red-633nm). Co-expression of insulin/SUR1 and insulin/Kir6.2 was analysed visually (green x red→yellow) and through correlation Pearson’s coefficient(PC) that estimates the goodness of rate association of the two fluorochromes. PC was compared among cases and controls using a nonparametric method (Mann-Whitney). Results: Visual observation clearly revealed the presence of Kir6.2 and SUR1 in a granular cytoplasmic pattern in the ß-cells of cases and controls. Nevertheless, overlap of insulin/Kir6.2 and insulin/SUR1 seemed to be more constant and uniformin controls than in CHI cases, confirmed by the analysis through PC showing a statistically significant decrease in Kir6.2 ( P = 0.0084) and SUR1 ( P = 0.041) in the ß-cells of CHI patients. Conclusions: This is the first demonstration of K ATP channels subunits Kir6.2 and SUR1 in the pancreatic ß-cells of CHI patients using an in situ method. Our results show that both subunits were present in the ß-cells, but are under-expressed in CHI patients compared to controls, adding a new information to this rare condition with a complex pathogenesis.conferenceObject Analysis of the Expression of Toll-like Receptors 2 and 4 in Keratinocytes of Patients with Extensive Dermatophytosis due Trichophyton rubrum(2012) CRIADO, P. R.; OLIVEIRA, C. B.; VASCONCELLOS, C.; VALENTE, N. Y. S.; SOTTO, M. N.; LUIZ, F. Guedes; BELDA JUNIOR, W.Rationale There are few studies to concern the role of innate immune response in dermatophytosis, so we conducted an investigation to define the involvement of TLRs in the course of tinea corporis by T. rubrum. Methods We allocated 14 patients without primary or secondary immunosuppression with extensive dermatophytosis, defined as the ringworm on at least 3 body segments of the same patient. In each patient the skin were biopsied from the active edge of the tinea and normal skin distant at least 4 cm of the lesion. Other skin fragments (control skin) were obtained from cosmetic surgery and without tinea. We use immunohistochemical staining with antibodies for antigens TLR 2 and 4. Images were analyzed in Image Pro Plus program. Results The Wilcoxon Signed Ranks Test showed: (i) regarding the expression of TLR2 of patients with tinea, found on the skin surface, average percentage of the marked area of 24.36 (1-76) in skin with tinea and 39.77 (9-84) in normal skin, p 0.043; (ii) analysis of TLR4 expression in the epidermis of patients with tinea met index higher optical density in normal skin than in skin with tinea, average 111.21 (99.44 to 134.34) and 104.50 (97.76 to 113.82), respectively, p 0.028. Conclusions We found a lower expression of TLR2 and 4 in the skin with tinea compared to healthy skin of the same patients with extensive dermatophytosis, as well as a tendency toward higher expression of TLR2 in the healthy peripheral skin, which could explain the spread in extension, in these cases of tinea.