Navegando por Palavras-chave "immunofluorescence"
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- ItemAcesso aberto (Open Access)Comparison of a rapid cytomegalovirus pp65 antigenemia assay revealed by immunofluorescence to an in-house assay revealed by immunoperoxidase for diagnosis in solid organ transplant recipient patients(Brazilian Society of Infectious Diseases, 2010-06-01) Carraro, Emerson [UNIFESP]; Pasternak, Jacyr [UNIFESP]; Perosa, Ana Helena Sitta [UNIFESP]; Siqueira, Itacy [UNIFESP]; Martino, Marines Dalla Vale [UNIFESP]; Hospital Albert Einstein Clinical Laboratory Microbiology Section; Universidade Federal de São Paulo (UNIFESP)Cytomegalovirus (CMV) antigenemia is still one of the two major assays available for diagnosis and monitoring of CMV infections. A commercial rapid test recently available in Brazil for quantification of human cytomegalovirus pp65 antigenemia revealed by immunofluorescence technique was compared with the original in-house method revealed by immunoperoxidase in patients receiving solid organ transplants. Of 80 blood samples tested for CMV antigenemia, 34 (42.5%) were positive: commercial assay detected 33 (97%) and in-house assay detected 20 (58.8%) samples. The numbers of positive cells in the two assays were different, with a median of 4.5 and 12 positive cells obtained by in-house and commercial kit, respectively. Discrepancies between assays occurred in 15 specimens from patients with low-grade antigenemia (median 6 positive cells). The assay-time was reduced in approximately 50% compared to in-house methodology. In conclusion, besides comparable results obtained for both assays, the commercial antigenemia assay provides more rapid and sensitive results.
- ItemSomente MetadadadosEffect of Collagen Cross-linking in Stromal Fibril Organization in Edematous Human Corneas(Lippincott Williams & Wilkins, 2010-07-01) Bottos, Katia Mantovani [UNIFESP]; Hofling-Lima, Ana Luisa [UNIFESP]; Barbosa, Manuela C. [UNIFESP]; Barbosa, Jose Bonifacio [UNIFESP]; Dreyfuss, Juliana L. [UNIFESP]; Schor, Paulo [UNIFESP]; Nader, Helena B. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Purpose: To assess structural stromal modifications after riboflavin and UV-A exposure in edematous human corneas.Method: Fourteen eyes with corneal edema were enrolled in the study. in the cross-linking (CXL) group, 7 corneal buttons were obtained from 6 patients who underwent penetrating keratoplasty (PK) 7-90 days after the CXL treatment. the control group was composed of 7 corneal buttons with bullous keratopathy. After the PK, stromal modifications were investigated using immunofluorescence in all corneal grafts. All patients had at least 3 months of corneal edema and were in the eye bank list waiting for keratoplasty.Results: All corneas in the treated group showed a pronounced lamellar zone of collagen fibers highly organized in the anterior stroma, but there was not complete homogeneity between the samples. Corneas with advanced disease and stromal fibrosis were less compacted than ones with mild disease severity. Similarly, those ones that underwent PK 3 months after CXL also showed a decreased effect compared with those with a reduced time between the CXL and the PK. DAPI staining demonstrated a complete fragmentation of keratocytes nuclei in the anterior stroma in all treated corneas, which were absent in the control group.Conclusions: Our study showed an immediate effect of CXL with a limited long-term sustainability. Cross-linked corneas had a pronounced anterior zone of organized collagen fibers. Even the treated corneas with advanced bullous keratopathy and stromal fibrosis had histological evidence of collagen fibers organization, but this effect seems to be decreased compared with corneas in initial stages of the disease.
- ItemAcesso aberto (Open Access)II Consenso Brasileiro de Fator Antinuclear em Células HEp-2: Definitions for standardization of autoantibody testing against the nucleus (ANA HEp-2), nucleolus, cytoplasm and mitotic apparatus, as wel as its clinical associations(Sociedade Brasileira de Reumatologia, 2003-06-01) Dellavance, Alessandra [UNIFESP]; Gabriel Júnior, Alexandre [UNIFESP]; Cintra, Alice Friedenberg de Ulhôa; Ximenes, Antônio Carlos; Nuccitelli, Barbara; Taliberti, Ben Hur; Moreira, Caio; Von Mühlen, Carlos Alberto; Bichara, Carlos David Araújo; Santos, Cláudio Henrique Ramos dos; Yano, Cristiane Martinez; Mangueira, Cristóvão Luis Pitangueiras; Carvalho, Darlene Gonçalves; Bonfá, Eloisa Silva Dutra de Oliveira; Doi, Elvira M.; Guimarães, Fabiana Nunes de Carvalho; Araújo, Flávia Ikeda e; Mundim, Hugo Mendonça; Rego, Jozelia; Vieira, Lisiane Ericonio dos Anjos; Poli, Luciana; Andrade, Luiz Eduardo Coelho [UNIFESP]; Callado, Maria Roseli; Mesquita, Mauro Meira; Sugiyama, Mitiko; Slhessarenko, Natasha; Silva, Nilzio Antônio da; Carballo, Orlando Gabriel; Leser, Paulo Guilherme [UNIFESP]; Francescantonio, Paulo Luiz Carvalho; Jarach, Renata; Xavier, Ricardo Machado; Levy, Roger Abramino; Neves, Suzane Pretti Figueiredo; Cruvinel, Wilson de Melo [UNIFESP]; Santos, Wilton Silva dos [UNIFESP]; Universidade de São Paulo (USP); Universidade Federal de São Paulo (UNIFESP); Bio-Rad Laboratório Brasil; Hospital Geral de Goiânia; Biomédica; Universidade Federal de Uberlândia; UFMG HC; PUCRS HC; New Life Produtos Hospitalares; Universidade Católica de Goiás; Patologista Clínica; Frischmann Aisengart Unidad Inmunología; Universidade Católica de Goiás Laboratório de Auto-Imunidade; Exame Medicina Laboratorial; FMUFG HC Laboratório de Imuno-Reumatologia e HLA; Lab. Santa Luzia; Medivax/Bion; HSPE/SP; Universidade Católica de Goiás Laboratório da Área de Saúde; Farmacêutica-Bioquímica; Univ. Fed. Mato Grosso; FMUFG Serviço de Reumatologia; Hospital Durand Unidad Inmunología; Laboratório Clínico; UFRGS HCPA Serviço de Reumatologia; UERJ FCM; UFMG FM; Hospital Universitário de Brasília Laboratório de ReumatologiaOBJECTIVE: The Second Brazilian Consensus on Antinuclear Antibodies (ANA) in HEp-2 Cells approved and extended the decision trees developed during the First Brazilian Consensus in order to also offer information about mixed patterns of fluorescence. METHODS: Since this test elicits reactions not only to nuclear autoimmune antigens but also to different cell compartments, new denominations for the test were approved. Results and CONCLUSIONS: These new denominations encompass variations on the autoantibody testing against the nucleus (ANA HEp-2), nucleolus, cytoplasm, and mitotic apparatus issue. Furthermore, major clinical associations were described for each immunofluorescent pattern, facilitating the interpretation of laboratory results in the clinical practice.
- ItemSomente MetadadadosSimultaneous isolation of platelet factor 4 and glycoprotein IIb-IIIa complex from rabbit platelets, and characterization of specific chicken antibodies to assay them(Elsevier B.V., 2004-01-01) Santoro, M. L.; Barbaro, K. C.; Rocha, TRF da; Torquato, RJS; Hirata, I. Y.; Sano-Martins, I. S.; Inst Butantan; Pro Sangue Fdn; Universidade Federal de São Paulo (UNIFESP)Rabbits are frequently used as models for studying coagulation and platelet disorders. However, few reports on literature have dealt with the purification and characterization of rabbit platelet proteins. Herein a protocol for the simultaneous purification of rabbit platelet factor 4 (PF4) and platelet glycoprotein IIb-IIIa (GPIIb-IIIa, integrin alpha(IIb)beta(3)) is described. Specific antibodies were raised in laying chicken, which were used for assaying PF4 by ELISA, and GPIIb-IIIa by direct immunofluorescence and flow cytometry. Furthermore, the binding of monoclonal antibodies specific for GPIIb-IIIa complex (P2), ligand-induced binding site of GPIIIa (LIBS1) and rabbit P-selectin (12A7), as well as of polyclonal IgY specific for rabbit GPIIb-IIIa, was compared in quiescent and thrombin-activated platelets. Polyclonal anti-rabbit PF4 IgY was a specific and sensitive probe that could be used for assaying PF4 in plasma samples. GPIIb-IIIa expression was increased in thrombin-activated platelets, as evaluated by flow cytometric analysis using P2 and polyclonal antibodies raised in chickens. Rabbit GPIIb-IIIa also exhibited a conformational modification that caused the appearance of ligand-induced binding sites. Increased P-selectin expression, used as a positive control, was also noticeable in thrombin-activated platelets. These data evidence that antibodies raised in laying chickens specific to rabbit PF4 and GPIIb-IIIa, as well as certain monoclonal antibodies specific for human GPIIb-IIIa, may be used for investigating rabbit platelet physiology. (C) 2003 Elsevier B.V. All rights reserved.