Eficacia del uso del calzado inestable en la calidad de vida de pacientes con linfedema de miembro inferior. Estudio piloto / Effectiveness of the use of unstable shoes in improving quality of life in lower limb lymphedema. A pilot study

Introducción. El linfedema se produce por una alteración en la circulación linfática. La bomba muscular y articular juegan un papel importante en dicha circulación, constituyendo mecanismos extrínsecos para favorecer el retorno, tanto venoso como linfático. El uso del calzado con suela inestable puede activar y fortalecer la musculatura de los miembros inferiores potenciando ambos mecanismos. El objetivo es evaluar la eficacia del uso del calzado con suela inestable en la calidad de vida en pacientes con linfedema de miembro inferior. Material y métodos. Se diseñó un ensayo clínico, controlado y aleatorizado, a doble ciego. Al tratamiento habitual del linfedema se añadió el uso de los zapatos con suela inestable (MBT®) al grupo experimental, para realizar las actividades diarias. Al grupo control se añadió el uso de zapatillas deportivas normales. Se reclutaron 21 sujetos. Se utilizó la encuesta EQ-5D de calidad de vida. Para el análisis comparativo se utilizó la t de Student. Resultados. Diecisiete sujetos completaron el estudio, de los cuales 10 usaron MBT® y 7 calzado deportivo convencional. Después de la intervención, ambos grupos mostraron un aumento de la puntuación media del cuestionario. Sin embargo, en el grupo que utilizó el calzado con suela inestable, el incremento fue de 18 puntos (p=0,003), frente a 11,4 (p=0,18) del grupo control. Conclusiones. Se puede considerar que el uso del calzado con suela inestable puede contribuir a mejorar la calidad de vida del paciente con linfedema del miembro inferior (AU)

Introduction. Lymphoedema is due to a lymph circulation disorder. Muscle and joint pumps are important extrinsic mechanisms of both lymphatic and venous return. The use of unstable shoes can activate and strengthen the muscles in the lower limbs, thus activating both mechanisms. The aim of this study was to evaluate the effectiveness of unstable shoes in improving quality of life in patients with lower limb lymphoedema. Material and methods. A double blind, randomised controlled trial was designed. In addition to their usual lymphoedema treatment, persons in the experimental group wore unstable shoes (MBT®) during their activities of daily living. The control group was given regular trainers. A total of 21 participants were recruited. Quality of life was measured using the EQ-5D questionnaire. Student's t- test was used for comparisons between groups. Results. Seventeen participants completed the study, of whom 10 used unstable shoes and 7 used another type of trainers. After the intervention, both groups showed an increase in mean quality of life scores. However, the mean score increased by 18 points in the group wearing MBT shoes (p=.003) and by 11.4 points in the control group (p=.18). Conclusions. The results of this study show that this kind of footwear may help to enhance quality of life in people with lower limb lymphoedema (AU)

Glycosylation and sorting pathways of lysosomal enzymes in mussel digestive cells.

Our aim was to contribute to the understanding of the synthesis, maturation and activation of lysosomal enzymes in an invertebrate cellular model: the endo-lysosomal system (ELS) of mussel digestive cells. The activities of 5'-nucleotidase (AMPase), arylsulphatase (ASase) and acid phosphatase (AcPase), which are transported towards acidic compartments as membrane proteins, were localised by enzyme cytochemistry. AcPase activity was found within large heterolysosomes and residual bodies. ASase was located in endosomes, endolysosomes and heterolysosomes. AcPase and ASase activities were recorded within small vesicles and cisterns of the trans-Golgi network. Conversely, AMPase activity was primarily found in microvilli and apical vesicles and, less conspicuously, in lysosomes and the cis-side of the Golgi and the cis-Golgi network (CGN). In order to understand the processes of synthesis and maturation of these lysosomal enzymes, selected glycoconjugates were localised after lectin cytochemistry. N-acetylglucosamine, mannose and fucose residues were almost ubiquitous in the ELS, as were galactose residues, which were apparently less abundant. N-acetylglucosamine residues occurred in the inner membrane co-localised with mannose residues within the lysosomal and pre-lysosomal acidic compartments. Based on these results, glycosylation and sorting pathways are proposed for both soluble and membrane enzymes. Unlike in mammalian cells, O-glycosylation is fully completed in the CGN, mannose addition in N-glycosylation extends beyond the CGN and galactose addition is fully achieved at the intermediate side. Sorting of soluble lysosomal enzymes, as in crustaceans, is mediated by the indirect transport of membrane-linked proteins with GlcNAc1-P6Man residues that are removed in endolysosomes and heterolysosomes.

Membrane flow through the Golgi apparatus: specific disassembly of the cis-Golgi network by ATP depletion.

Incubation of NRK cells for 30 to 45 minutes with 50 mM 2-deoxy-D-glucose (DOG) in glucose and pyruvate-free medium results in depletion of the cellular ATP pool and in specific disassembly of the cis-Golgi network (CGN), with the stack of Golgi cisternae (SGC) and the trans-Golgi network (TGN) remaining intact and sensitive to BFA. The disassembly of the CGN is mediated by long tubular structures extending outwards from the Golgi complex and involves microtubules. Upon removal of DOG and addition of glucose and pyruvate to the culture medium, the morphology of the CGN is slowly reestablished. Reconstruction of the CGN involves COPI/COPII-positive vesicles that resume the transport of proteins and in particular of CGN membrane proteins out of the ER. Exit of CGN membrane proteins from the ER is insensitive to BFA. In cells pretreated with nocodazole, the CGN membrane proteins are transported to the vicinity of the SGC fragments dispersed throughout the cytoplasm. Ultrastructural studies of cells engaged in the reconstruction of the CGN revealed that the CGN cisterna emerge as tubular structures extending from 0.2-0.3 microm uncoated vesicles prior to their organization on the cis-side of the SGC.

Analysis of the distribution of glycoconjugates in the digestive gland of the bivalve mollusc Mytilus galloprovincialis by conventional and lectin histochemistry.

We examined the distribution and pattern of reactivity of a panel of 16 lectins in the digestive gland of the bivalve mollusc Mytilus galloprovincialis at the light-microscopic level. Various chemical treatments were applied in combination with lectins to differentiate between N- and O-linked oligosaccharides. Several control reactions were carried out, including replacement of lectins by buffer and incubation with their specific competitive inhibitors. Some lectins reacted selectively with particular cell types, thus revealing a cell-specific glycoconjugate distribution pattern which is possibly related to the metabolic role of each cell type in the digestive gland. Glycoconjugates containing glucosamine, mannose, and sulfated galactose were associated with the endolysosomal system of digestive cells. These glycoconjugates were also found in small vesicles randomly distributed in the cytoplasm of adipogranular cells in the connective tissue. However galactosamine residues appeared to be associated mainly with basophilic cells. Fucose residues did not exhibit a cell-specific distribution and appeared in small amounts homogeneously distributed throughout the digestive gland tissue. Conventional histochemical reactions for carbohydrate detection revealed moderate amounts of periodic-acid-Schiff-positive, neutral carbohydrates widely distributed in digestive and connective tissues. Among the acid carbohydrates, most cell types contained complex sulfated carbohydrates, but not carboxylated ones; this agreed well with the complete lack of sialic acid residues in all cell types studied, as observed by lectin histochemistry.

Isolation and morphofunctional characterization of mussel digestive gland cells in vitro.

The aim of the present study was to isolate and characterize digestive gland cells of the bivalve mollusc Mytilus galloprovincialis Lmk. The digestive gland of bivalves is a complex organ composed of digestive and connective tissues but it is also invaded by the reproductive tissue as gametogenesis proceeds. The digestive tissue is comprised of stomach and intestinal epithelial cells, ciliated and non-ciliated duct cells, digestive cells and basophilic cells. the last two cell types are found lining the epithelium of the blind-ending digestive tubules and are the main focus of this study. Two different approaches were assayed for cell isolation, i.e., explant culture techniques and mechanical plus enzymatical digestion techniques. Cell viability was tested by trypan blue exclusion, neutral red uptake and ultrastructural analysis. The explant cultures were often contaminated with bacteria and spermatozoa and, moreover, cells migrating out of the explants possibly corresponded to hemocytes and not to digestive tissue cells. Mechanical plus enzymatical digestion with collagenase was concluded to be the method of choice for digestive cell isolation, with a percentage of about 30% of isolated cells corresponding to digestive cells. The ultrastructure of digestive cells isolated with the latter procedure closely resembled that of mussel digestive cells in vivo.

Ultrastructural and cytochemical study of the Golgi complex of molluscan (Mytilus galloprovincialis) digestive cells

The present study has been performed to investigate the ultrastructure and subcellular distribution of several marker enzymes within the Golgi complex of digestive cells of the mussel, Mytilus galloprovincialis. This cell type is involved in the intracellular digestion of food and contains a well-developed lysosomal-vacuolar system and a characteristic cup-shaped Golgi complex. The number and degree of compaction of the tubulo-filamentous structures extending across the distended regions of the cisternae, the number of Golgi stacks per cell and the number of cisternae per stack vary with season and nutritional status of the animals. The degree of compaction of the tubulo-filamentous structures defines the cis- and trans-portions of the Golgi apparatus, the less compacted structures being associated with the cis-portion. Acid phosphatase and arylsulphatase activities are located within some tubules and vesicles in the trans-Golgi network and in some membrane-bounded organelles comprising the lysosomal-vacuolar system. The tubulo-filamentous structures of the dilated rims are reactive for arylsulphatase activity only when located in the trans-portion. Osmium impregnation techniques label all the cisternae of the Golgi complex. No nicotinamide adenine dinucleotide phosphatase activity is present in Golgi bodies or other associated structures in digestive cells. Thiamine pyrophosphatase activity only rarely occurs in some elongate tubules that are interpreted as belonging to trans-cisternae.