Tubo de ácido poliglicólico e GM1 na regeneração de nervos periféricos

INTRODUÇÃO: A auto-enxertia de nervo é considerada tratamento de escolha nas grandes perdas de tecido neural que não permitam a reparação através de anastomose primária. Nesses casos, o tubo sintético à base de ácido poliglicólico é uma alternativa para enxertia de nervo. Por outro lado, muitos estudos têm enfatizado a importância dos fatores neurotróficos na regeneração neural: o monossialotetraesosilgangliosídeo (GM1), um dos principais glicoesfingolípides do tecido nervoso de mamíferos, é tido como potencializador dos efeitos desses fatores. OBJETIVO: Comparar, em ratos, o grau de regeneração neural, utilizando análise histológica, contagem do número de axônios mielinizados regenerados e análise funcional com a utilização do neurotubo e do GM1. MÉTODOS: Essa avaliação foi obtida com a interposição de enxerto autógeno (grupo A), tubo de ácido poliglicólico (grupo B) e da associação do tubo de ácido poliglicólico à administração de GM1 (grupo C) em defeitos de 5 mm no nervo ciático. RESULTADOS: Foi observada formação de neuroma apenas no grupo A. Os grupos A e C apresentaram padrões histológicos semelhantes, exceto que os axônios regenerados do grupo C apresentavam-se mais organizados e mielinizados que o grupo A. CONCLUSÃO: Na recuperação funcional, não houve diferença estatisticamente significativa entre os três grupos, a despeito das diferenças histológicas qualitativas e quantitativas verificadas.

Potentializing the effects of GM1 by hyperbaric oxygen therapy in acute experimental spinal cord lesion in rats

Study design: Experimental, controlled, animal study. Objectives: To evaluate the effect of GM1 ganglioside, hyperbaric oxygen and both in combination, in the treatment of experimental spinal cord lesions in rats. Setting: Brazil. Methods: Thirty-two Wistar rats with spinal cord lesions were divided into four groups: one group received GM1 ganglioside, one was submitted to hyperbaric oxygen therapy (HBOT), the third received both treatments and the fourth received no treatment (control). Results: There were no significant differences between the groups in the histological analysis, for any of the variables (necrosis, hemorrhage, hyperemia, cystic degeneration, P>0.06). Neither were there any significant differences in the comparison of left and right sides in the functional tests (P>0.06 for all). No significant differences were found in the locomotor ratings, in the comparison of groups at 2, 7, 21 and 28 days after the surgical procedure. However, in the evaluation on day 14, group 3, which received the combined therapy, showed a significantly higher Basso Beattie and Bresnahan score than the other groups (P = 0.015). Conclusion: The therapeutic effect of GM1 in locomotor evaluation of rats submitted to spinal cord lesion is anticipated by HBOT. Spinal Cord (2010) 48, 808-813; doi:10.1038/sc.2010.37; published online 27 April 2010

POLYGLYCOLIC ACID TUBE ASSOCIATED WITH GM1 IN REGENERATION OF PERIPHERAL NERVES

Introduction: Nerve allografting is regarded as a treatment of choice in large neural tissue losses preventing repair by primary anastomosis. In these cases, a synthetic polyglycolic acid tube is an alternative for nerve grafting. On the other hand, several studies have emphasized the importance of neurotrophic factors on neural regeneration, including substances with potential to optimize neural regeneration, especially the GM1, an neurotrophic enhancer factor. Objective: to compare, in rats, the neural regeneration degree using histological analysis, regenerated myelinized axons count, and functional analysis with the use of neurotube and GM1. Methods: This assessment was performed by interposing allograft (group A), polyglycolic acid tube (group B) and polyglycolic acid tube associated to GM1 (group C) on 5-mm sciatic nerve defects. Results: Neuroma formation was found only on group A. Groups A and C showed similar histological patterns, except for the regenerated axons on group C, which were shown to be better organized and myelinized than in group A. Conclusion: on functional recovery, no statistically significant difference was found for the three groups, despite of qualitative and quantitative histological differences found.

GD1b-Derived Gangliosides Modulate Fc epsilon RI Endocytosis in Mast Cells

The role of the mast cell-specific gangliosides in the modulation of the endocytic pathway of Fc epsilon RI was investigated in RBL-2H3 cells and in the ganglioside-deficient cell lines, E5 and D1. MAb BC4, which binds to the alpha subunit of Fc epsilon RI, was used in the analysis of receptor internalization. After incubation with BC4-FITC for 30 min, endocytic vesicles in RBL-2H3 and E5 cells were dispersed in the cytoplasm. After 1 hr, the endocytic vesicles of the RBL-2H3 cells had fused and formed clusters, whereas in the E5 cells, the fusion was slower. In contrast, in D1 cells, the endocytic vesicles were smaller and remained close to the plasma membrane even after 3 hr of incubation. When incubated with BC4-FITC and subsequently imunolabeled for markers of various endocytic compartments, a defect in the endocytic pathway in the E5 and D1 cells became evident. In the D1 cells, this defect was observed at the initial steps of endocytosis. Therefore, the ganglioside derivatives from GD1b are important in the endocytosis of Fc epsilon RI in mast cells. Because gangliosides may play a role in mast cell-related disease processes, they provide an attractive target for drug therapy and diagnosis. (J Histochem Cytochem 59:428-440, 2011)

Genetic diversity of heat-labile toxin expressed by enterotoxigenic Escherichia coli strains isolated from humans

The natural diversity of the eft operons, encoding the heat-labile toxin LT-I (LT), carried by enterotoxigenic Escherichia coli (ETEC) strains isolated from humans was investigated. For many years, LT was supposed to be represented by a rather conserved toxin, and one derivative, produced by the reference H10407 strain, was intensively studied either as a virulence factor or as a vaccine adjuvant. Amplicons encompassing the two LT-encoding genes (eltA and eltB) of 51 human-derived ETEC strains, either LT+ (25 strains) only or LT+/ST+ (26 strains), isolated from asymptomatic (24 strains) or diarrheic (27 strains) subjects, were subjected to restriction fragment length polymorphism (RFLP) analysis and DNA sequencing. Seven polymorphic RFLP types of the H10407 strain were detected with six (BsaI, DdeI, HhaI, HincII, HphI, and MspI) restriction enzymes. Additionally, the single-nucleotide polymorphic analysis revealed 50 base changes in the eft operon, including 21 polymorphic sites at eltA and 9 at eltB. Based on the deduced amino acid sequences, 16 LT types were identified, including LT1, expressed by the H10407 strain and 23 other strains belonging to seven different serotypes, and LT2, expressed by 11 strains of six different serotypes. In vitro experiments carried out with purified toxins indicated that no significant differences in GM1-binding affinity could be detected among LT1, LT2, and LT4. However, LT4, but not other toxin types, showed reduced toxic activities measured either in vitro with cultured cells (Y-1 cells) or in vivo in rabbit ligated ileal loops. Collectively, these results indicate that the natural diversity of LTs produced by wild-type ETEC strains isolated from human hosts is considerably larger than previously assumed and may impact the pathogeneses of the strains and the epidemiology of the disease.

The specificity of frutalin lectin using biomembrane models

Frutalin is a homotetrameric alpha-D-galactose (D-Gal)-binding lectin that activates natural killer cells in vitro and promotes leukocyte migration in vivo. Because lectins are potent lymphocyte stimulators, understanding the interactions that occur between them and cell surfaces can help to the action mechanisms involved in this process. In this paper, we present a detailed investigation of the interactions of frutalin with phospho- and glycolipids using Langmuir monolayers as biomembrane models. The results confirm the specificity of frutalin for D-Gal attached to a biomembrane. Adsorption of frutalin was more efficient for the galactose polar head lipids, in contrast to the one for sulfated galactose, in which a lag time is observed, indicating a rearrangement of the monolayer to incorporate the protein. Regarding ganglioside GM1 monolayers, lower quantities of the protein were adsorbed, probably due to the farther apart position of D-galactose from the interface. Binary mixtures containing galactocerebroside revealed small domains formed at high lipid packing in the presence of frutalin, suggesting that lectin induces the clusterization and the forming of domains in vitro, which may be a form of receptor internalization. This is the first experimental evidence of such lectin effect, and it may be useful to understand the mechanism of action of lectins at the molecular level. (C) 2010 Elsevier B.V. All rights reserved.