A comparative analysis of structure and spatial distribution of decorin inhuman leiomyoma and normal myometrium

Leiomyoma is a benign smooth muscle tumor of the uterus that affects many women in active reproductive life. It is composed by bundles of smooth muscle cells surrounded by extracellular matrix. We have recently shown that the glycosylation of extracellular matrix proteoglycans is modified in leiomyoma: increased amounts of galactosaminoglycans with structural modifications are present. the data here presented show that decorin is present in both normal myometrium and leiomyoma but tumoral decorin is glycosylated with longer galactosaminoglycan side chains. Furthermore, these chains contain a higher ratio D-glucuronate/L-iduronate, as compared to normal tissue. To determine if these changes in proteoglycan glycosylation correlates with modifications in the extracellular matrix organization, we compared the general structural architecture of leiomyoma to normal myometrium. By histochemical and immunofluorescence methods, we found a reorganization of muscle fibers and extracellular matrix, with changes in the distribution of glycoproteins, proteoglycans, and collagen. Thin reticular fibers, possibly composed by types I and III collagen, were replaced by thick fibers, possibly richer in type I collagen. Type I collagen colocalized with decorin both in leiomyoma and normal myometrium, in contrast to type IV collagen that did not. the relative amount of decorin was increased and the distribution of decorin and collagen was totally modified in the tumor, as compared to the normal myometrium. These findings reveal that not only decorin structure is modified in leiomyoma but also the tissue architecture changed, especially concerning extracellular matrix. (C) 2002 Elsevier Science B.V. All rights reserved.UNIFESP EPM, Dept Bioquim, Disciplina Biol Mol, BR-04044020 São Paulo, SP, BrazilUniv São Paulo, Inst Matemat & Estatist, Dept Ciencias Computacao, São Paulo, SP, BrazilUNIFESP EPM, Dept Bioquim, Disciplina Biol Mol, BR-04044020 São Paulo, SP, BrazilWeb of Scienc

Fucan inhibits Chinese hamster ovary cell (CHO) adhesion to fibronectin by binding to the extracellular matrix

In recent years, sulfated fucans have emerged as an important class of natural biopolymers. in this study, the anti-adhesive activity of a fucan from the brown seaweed Spatoglossum schroederi was analyzed using tumorigenic cells: wild-type Chinese hamster ovary cells (CHO-K1) and the mutant type deficient in xylosyltransferase (CHO-745). Fibronectin (FN) was used as substrate for cell attachment. for both cell types, this fucan has shown a dose-dependent anti-adhesive effect, reaching saturation at around 400 mu g/mL. This effect was abolished by desulfation of the fucan. in addition, this polymer exhibited the highest inhibitory effect in comparison to other sulfated polysaccharides. the fucan was biotinylated and used as a probe to identify its action sites. Biotinylated fucan was detected in the extracellular matrix environment by confocal microscopy and flow cytometric analysis, but not at the cell surface. the results suggest that the fucan shows anti-adhesive activity by binding directly to FN, and blocking FN sites that are recognized by cell surface ligands, possibly the integrin family.Universidade Federal de São Paulo, Dept Bioquim, Escola Paulista Med, BR-04044020 São Paulo, BrazilUniv Fed Rio Grande Norte, Lab Biotecnol Polimeros Nat, BIOPOL, Dept Bioquim, BR-59072970 Natal, RN, BrazilUniv Fed Parana, Dept Biol Celular, BR-80060000 Curitiba, Parana, BrazilUniversidade Federal de São Paulo, Dept Bioquim, Escola Paulista Med, BR-04044020 São Paulo, BrazilWeb of Scienc

Structural and ultrastructural description of the venom gland of Loxosceles intermedia (brown spider)

The brown spider, genus Loxosceles, is becoming of great medical importance, with envenomation (Loxoscelism) occurring throughout the world. the biological activities of the brown spider venom usually include dermonecrotic lesions at the bite site accompanied by hemolytic and haemorrhagic effects and also by renal failure. the objective of the present study was to describe the histology of the venom gland of L. intermedia using glands from adult spiders which were investigated by light microscopy, using immunohistochemical and staining methods, by transmission electron microscopy, and by scanning electron microscopy. the organization of the venom gland of Loxosceles intermedia follows the general architecture of spiders' venom glands. Using light microscopy and transmission electron microscopy we observed that the venom glands of L. intermedia present two layers of striated muscle fibers, an external layer and an internal layer in touch with an extracellular matrix which is a basement membrane structure and a fibrillar collagen matrix separating the muscular region from epithelial cells of the venom gland. Muscle cells are multinucleated, with nuclei peripherally placed and their cytoplasm rich in sarcoplasmic reticulum, myofibrills and continuous Z lines. By using scanning electron microscopy we can detect muscular cells from external layer as branching cells. Epithelial cells have their cytosol extremely rich in rough endoplasmic reticulum, mitochondria collection, Golgi apparatus, interdigitating membranes and secretory vesicles that ultimately accumulate the venom, a complex protein mixture. (C) 1999 Elsevier B.V. All rights reserved.Univ Fed Parana, Dept Cell Biol, BR-80060000 Curitiba, Parana, BrazilUniv Fed Parana, Dept Physiol, BR-80060000 Curitiba, Parana, BrazilUniversidade Federal de São Paulo, Dept Biochem, São Paulo, BrazilUniv Fed Parana, Cent Lab, LACTEC, COPEL, BR-80060000 Curitiba, Parana, BrazilUniversidade Federal de São Paulo, Dept Biochem, São Paulo, BrazilWeb of Scienc

In vivo and in vitro cytotoxicity of brown spider venom for blood vessel endothelial cells

The effect of brown spider (Loxosceles intermedia) venom on endothelial cells was investigated in vivo and in vitro. Morphological and ultrastructural observations by light microscopy and transmission electron microscopy showed that the venom acts in vivo upon vessel endothelial cells of rabbits that were intradermally injected, evoking vessel instability, cytoplasmic endothelial cell vacuolization, and blebs. Likewise, treatment of rabbit endothelial cells in culture with the venom led to loss of adhesion of the cells to the substrate. Endothelial cells in culture were metabolically radiolabeled with sodium [S-35]- sulfate and the sulfated compounds (proteoglycans and sulfated proteins) from medium, cell surface, and extracellular matrix (ECM) were analyzed. Agarose gel electrophoresis and SDS-PAGE showed that the venom is active on the ECM and on cell surface proteoglycans, shedding these molecules into the culture medium. in addition, when purified heparan sulfate proteoglycan (HSPG) and purified laminin-entactin (LN/ET) complex were incubated with the venom we observed a partial degradation of the protein core of HSPG as well as the hydrolysis of entactin. the above results suggest that the L. intermedia venom has a deleterious effect on the endothelium of vessels both in vivo and in culture, removing important constituents such as HSPG and entactin that are involved in the adhesion of endothelial cells and of subendothelial ECM organization. (C) 2001 Elsevier B.V. All rights reserved.Univ Fed Parana, Dept Cell Biol, BR-81531990 Curitiba, Parana, BrazilUniv Fed Parana, Dept Physiol, BR-80060000 Curitiba, Parana, BrazilUniversidade Federal de São Paulo, Dept Biochem, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biochem, São Paulo, BrazilWeb of Scienc

Structural and hemostatic activities of a sulfated galactofucan from the brown alga Spatoglossum schroederi - An ideal antithrombotic agent?

The brown alga Spatoglossum schroederi contains three fractions of sulfated polysaccharides. One of them was purified by acetone fractionation, ion exchange, and molecular sieving chromatography. It has a molecular size of 21.5 kDa and contains fucose, xylose, galactose, and sulfate in a molar ratio of 1.0:0.5:2.0:2.0 and contains trace amounts of glucuronic acid. Chemical analyses, methylation studies, and NMR spectroscopy showed that the polysaccharide has a unique structure, composed of a central core formed mainly by 4-linked beta-galactose units, partially sulfated at the 3-O position. Approximately 25% of these units contain branches of oligosaccharides (mostly tetrasaccharides) composed of 3-sulfated, 4-linked alpha-fucose and one or two nonsulfated, 4-linked beta-xylose units at the reducing and nonreducing end, respectively. This sulfated galacto-fucan showed no anticoagulant activity on several in vitro assays. Nevertheless, it had a potent antithrombotic activity on an animal model of experimental venous thrombosis. This effect is time-dependent, reaching the maximum 8 h after its administration compared with the more transient action of heparin. the effect was not observed with the desulfated molecule. Furthermore, the sulfated galactofucan was 2-fold more potent than heparin in stimulating the synthesis of an antithrombotic heparan sulfate by endothelial cells. Again, this action was also abolished by desulfation of the polysaccharide. Because this sulfated galactofucan has no anticoagulant activity but strongly stimulates the synthesis of heparan sulfate by endothelial cells, we suggested that this last effect may be related to the in vivo antithrombotic activity of this polysaccharide. in this case the highly sulfated heparan sulfate produced by the endothelial cells is in fact the antithrombotic agent. Our results suggested that this sulfated galactofucan may have a potential application as an antithrombotic drug.Universidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, BrazilUniv Fed Rio Grande Norte, Dept Bioquim, Lab Biotecnol Polimeros Nat BIOPOL, BR-59072970 Natal, RN, BrazilUniv Fed Parana, Dept Biol Celular, BR-83531990 Curitiba, Parana, BrazilUniv Fed Rio de Janeiro, Inst Bioquim Med, BR-21941590 Rio de Janeiro, BrazilUniv Fed Rio de Janeiro, Univ Hosp, BR-21941590 Rio de Janeiro, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, BrazilWeb of Scienc

Fluorescence properties of curcumin-loaded nanoparticles for cell tracking

Bassam Felipe Mogharbel,1 Julio Cesar Francisco,1 Ana Carolina Irioda,1 Dilcele Silva Moreira Dziedzic,1 Priscila Elias Ferreira,1 Daiany de Souza,1 Carolina Maria Costa de Oliveira Souza,1 Nelson Bergonse Neto,2 Luiz Cesar Guarita-Souza,2 Celia Regina Cavichiolo Franco,3 Celso Vataru Nakamura,4 Vanessa Kaplum,4 Letícia Mazzarino,5 Elenara Lemos-Senna,6 Redouane Borsali,7 Paula A Soto,8 Patricia Setton-Avruj,8 Eltyeb Abdelwahid,9 Katherine Athayde Teixeira de Carvalho1 1Cell Therapy and Biotechnology in Regenerative Medicine Department, Pelé Pequeno Príncipe Institute, Child and Adolescent Health Research and Pequeno Príncipe Faculty, Curitiba, Paraná, Brazil; 2Institute of Biological and Health Sciences, Pontifical Catholic University of Paraná (PUCPR), Centro de Ciências Biológicas e da Saúde (CCBS), Curitiba, Brazil; 3Cell Biology Department, Federal University of Paraná, Curitiba, Paraná, Brazil; 4Department of Pharmaceutical Sciences, Universidade Estadual de Maringá, Maringá, Paraná, Brazil; 5Department of Pharmaceutical Sciences, NanoBioMat Laboratory, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil; 6Department of Pharmaceutical Sciences, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil; 7Centre de Recherches sur les Macromolécules Végétales (CERMAV), Centre National de la Recherche Scientifique (CNRS), University Grenoble Alpes, F-38000, Grenoble, France; 8Instituto de Química y Físicoquímica Biológica (IQUIFIB), Departament of Química Biológica, Facultad de Farmacia y Bíoquímica, Universidad de Buenos Aires (UBA) Consejo nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentine; 9Feinberg School of Medicine, Feinberg Cardiovascular Research Institute, Northwestern University, Chicago, Il, USA Background: Posttransplant cell tracking, via stem cell labeling, is a crucial strategy for monitoring and maximizing benefits of cell-based therapies. The structures and functionalities of polysaccharides, proteins, and lipids allow their utilization in nanotechnology systems. Materials and methods: In the present study, we analyzed the potential benefit of curcumin-loaded nanoparticles (NPC) using Vero cells (in vitro) and NPC-labeled adipose-derived mesenchymal stem cells (NPC-ADMSCs) (in vivo) in myocardial infarction and sciatic nerve crush preclinical models. Thereafter, transplantation, histological examination, real time imaging, and assessment of tissue regeneration were done. Results: Transplanted NPC-ADMSCs were clearly identified and revealed potential benefit when used in cell tracking. Conclusion: This approach may have broad applications in modeling labeled transplanted cells and in developing improved stem cell therapeutic strategies. Keywords: mesenchymal stem cells, transplantation, cell marking, myocardium infarction, sciatic nerve crus