Apical hypertrophic cardiomyopathy : a review

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

(Table 2) Rate of reduction in cell size (apical length) and growth rate for seven Fragilariopsis kerguelensis strains sampled during POLARSTERN cruise ANT-XXI/3

The planktonic diatom Fragilariopsis kerguelensis plays an important role in the biogeochemical cycles of the Southern Ocean, where remains of its frustules form the largest deposit of biogenic silica anywhere in the world. We assessed the genetic identity of 26 strains, from cells collected at various sites in the Southern Ocean, using three molecular markers, LSU and ITS rDNA and rbcL. The LSU sequences were identical among the tested strains, ITS sequences were highly similar, and only one base pair difference was detected among the rbcL sequences. These results, together with a large number of successful mating experiments demonstrated that the strains belong to a single biological species. We investigated the mating system and life cycle traits of F. kerguelensis. Cell size diminished gradually in clonal strains. Gamete formation only occurred when strains of opposite mating type - within a cell size range of 7-36 µm - were mixed together. Two binucleate gametes were formed in each gametangium and gamete conjugation produced a zygote that had four nuclei and was surrounded by thin siliceous scales. Two out of the four nuclei subsequently degenerated and the zygote expanded to form an auxospore surrounded by a transverse and a longitudinal perizonium. Staining with the fluorochrome PDMPO provided for the first time a clear demonstration that the longitudinal perizonium is formed after auxospore expansion is complete. Initial cells produced within the mature auxospores were 78-101 µm in length. Various authors have shown that the average valve size of F. kerguelensis varies in sediment samples collected in regions and seasons with different primary production regimes and this parameter has thus been proposed as a biological proxy for palaeo-productivity. A better understanding of the life cycle of F. kerguelensis should help the design of future investigations aimed at testing the link between cell size distribution in the natural environment and the role that environmental factors might have in the regulation of population cell size.

Culture of apical meristems and embryonic shoots of picea abies --approach and techniques

Bibliography: p. 29-30.

Dominância política e paralisia executiva

Desde 2014 nossas análises têm enfatizado o vínculo entre a política e a economia para explicar a crise pela qual o país passa desde então. De um lado, a falta de consenso político tem impedido respostas mais fortes ao quadro de deterioração fiscal. Isso derrubou a confiança de empresários e consumidores, levando junto o nível de atividade e, em um círculo vicioso, as receitas tributárias, dessa forma agravando ainda mais o quadro fiscal. De outro lado, a deterioração econômica enfraqueceu a disposição dos políticos, a começar pelo próprio Executivo, de adotar medidas mais fortes na área fiscal, levando a um quadro de quase inércia na política econômica.

Carboxy terminus of glucose transporter 3 contains an apical membrane targeting domain

We previously demonstrated that distinct facilitative glucose transporter isoforms display differential sorting in polarized epithelial cells. In Madin-Darby canine kidney (MDCK) cells, glucose transporter 1 and 2 (GLUT1 and GLUT2) are localized to the basolateral cell surface whereas GLUTs 3 and 5 are targeted to the apical membrane. To explore the molecular mechanisms underlying this asymmetric distribution, we analyzed the targeting of chimeric glucose transporter proteins in MDCK cells. Replacement of the carboxy-terminal cytosolic tail of GLUT1, GLUT2, or GLUT4 with that from GLUT3 resulted in apical targeting. Conversely, a GLUT3 chimera containing the cytosolic carboxy terminus of GLUT2 was sorted to the basolateral membrane. These findings are not attributable to the presence of a basolateral signal in the tails of GLUTs 1, 2, and 4 because the basolateral targeting of GLUT1 was retained in a GLUT1 chimera containing the carboxy terminus of GLUT5. In addition, we were unable to demonstrate the presence of an autonomous basolateral sorting signal in the GLUT1 tail using the low-density lipoprotein receptor as a reporter. By examining the targeting of a series of more defined GLUT1/3 chimeras, we found evidence of an apical targeting signal involving residues 473 - 484 (DRSGKDGVMEMN) in the carboxy tail. We conclude that the targeting of GLUT3 to the apical cell surface in MDCK cells is regulated by a unique cytosolic sorting motif.

Specific modulation of airway epithelial tight junctions by apical application of an occludin peptide

Tight junctions are directly involved in regulating the passage of ions and macromolecules (gate functions) in epithelial and endothelial cells. The modulation of these gate functions to transiently regulate the paracellular permeability of large solutes and ions could increase the delivery of pharmacological agents or gene transfer vectors. To reduce the inflammatory responses caused by tight junction-regulating agents, alternative strategies directly targeting specific tight junction proteins could prove to be less toxic to airway epithelia. The apical delivery of peptides corresponding to the first extracellular loop of occludin to transiently modulate apical paracellular flux has been demonstrated in intestinal epithelia. We hypothesized that apical application of these occludin peptides could similarly modulate tight junction permeability in airway epithelia. Thus, we investigated the effects of apically applied occludin peptide on the paracellular permeability of molecular tracers and viral vectors in well differentiated human airway epithelial cells. The effects of occludin peptide on cellular toxicity, tight junction protein expression and localization, and membrane integrity were also assessed. Our data showed that apically applied occludin peptide significantly reduced transepithelial resistance in airway epithelia and altered tight junction permeability in a concentration-dependent manner. These alterations enhanced the paracellular flux of dextrans as well as gene transfer vectors. The occludin peptide redistributed occludin but did not alter the expression or distribution of ZO-1, claudin-1, or claudin-4. These data suggest that specific targeting of occludin could be a better-suited alternative strategy for tight junction modulation in airway epithelial cells compared with current agents that modulate tight junctions.

Retromer controls epithelial cell polarity by trafficking the apical determinant crumbs

The evolutionarily conserved apical determinant Crumbs (Crb) is essential for maintaining apicobasal polarity and integrity of many epithelial tissues [1]. Crb levels are crucial for cell polarity and homeostasis, yet strikingly little is known about its trafficking or the mechanism of its apical localization. Using a newly established, liposome-based system described here, we determined Crb to be an interaction partner and cargo of the retromer complex. Retromer is essential for the retrograde transport of numerous transmembrane proteins from endosomes to the trans-Golgi network (TGN) and is conserved between plants, fungi, and animals [2]. We show that loss of retromer function results in a substantial reduction of Crb in Drosophila larvae, wing discs, and the follicle epithelium. Moreover, loss of retromer phenocopies loss of crb by preventing apical localization of key polarity molecules, such as atypical protein kinase C (aPKC) and Par6 in the follicular epithelium, an effect that can be rescued by overexpression of Crb. Additionally, loss of retromer results in multilayering of the follicular epithelium, indicating that epithelial integrity is severely compromised. Our data reveal a mechanism for Crb trafficking by retromer that is vital for maintaining Crb levels and localization. We also show a novel function for retromer in maintaining epithelial cell polarity.

High permeability of the anionic form restricts accumulation of indomethacin by cultured gastric surface epithelial cells exposed to low apical pH

The 'ion-trapping' hypothesis suggests that the intracellular concentration of acidic non-steroidal anti-inflammatory drugs in gastric epithelial cells could be much higher than in the gastric lumen, and that such accumulation could contribute to their gastrotoxicity. Our aim was to examine the effect of the pH of the apical medium on the apical to basal transfer of the acidic drug indomethacin (pK a 4.5) across a gastric mucous epithelial cell monolayer, and to determine whether indomethacin accumulated in cells exposed to a low apical pH. Guinea-pig gastric mucous epithelial cells were grown on porous membrane culture inserts (Transwells®) for 72 h. Transfer and accumulation of [ 14C] indomethacin were assessed by scintillation counting. Transfer of [ 3H]mannitol and measurement of trans-epithelial electrical resistance were used to assess integrity of the monolayer. Distribution of [ 14C] urea was used to estimate the intracellular volume of the monolayer. The monolayer was not disrupted by exposure of the apical face to media of pH ≥ 3, or by indomethacin. Transfer of indomethacin (12 μM) to the basal medium increased with decreasing apical medium pH. The apparent permeability of the undissociated acid was estimated to be five times that of the anion. The intracellular concentration of indomethacin was respectively 5.3, 4.1 and 4.3 times that in the apical medium at pH 5.5, 4.5 and 3.0. In conclusion, this study represents the first direct demonstration that indomethacin accumulates in gastric epithelial cells exposed to low apical pH. However, accumulation of indomethacin was moderate and the predictions of the ion-trapping hypothesis were not met, probably due to the substantial permeability of anionic indomethacin across membranes. © 2006 Elsevier B.V. All rights reserved.

Apicectomías de dientes premolares y molares mandibulares en el conejo de raza neo zelandesa: estudio histológico usando distintos materiales de obturación apical

La apicectomía es una técnica de cirugía odontológica humana y veterinaria, que consiste en la amputación o resección de la porción más apical de la raíz dentaria y la retrobturación del ápice con algún material, con el objetivo de conservar el diente afectado de alguna patología apical. Se han usado gran variedad de materiales para la obturación, como amalgama, gutapercha, ionómero de vidrio, composite, MTA e hidróxido de calcio. En este estudio pretendemos comprobar la viabilidad de la técnica de la realización de la apicectomías en premolares y molares mandibulares, usando distintos materiales de retrobturación como son el iónomero de vidrio Ketac Cem μ® y el composite Tetric EvoFlow® en el conejo de experimentación de raza Neozelandesa. Realizamos las apicectomías de los dientes 407, 408 y 409 (3º premolar, 4º premolar y 1º molar inferior derecho), en una sola fase a diez animales. En el primer diente (407) solo se aplica un sellado del diente apicectomizado con hidróxido de calcio en polvo® y pasta Dycal® Dentin y no se coloca ningún material de obturación. En el segundo diente (408), se aplica un sellado con hidróxido de calcio en polvo® y pasta Dycal® Dentin y encima una retrobturación con ionómero de vidrio Ketac Cem μ® y en el tercer diente (409), se aplica un sellado con hidróxido de calcio en polvo® y pasta Dycal® Dentin y encima una retroturación con el composite Tetric EvoFlow®. A las 4 y 8 semanas se sacrificaron los animales y se realizó la toma de muestras. Estas fueron radiografiadas y posteriormente se realizaron cortes histológicos de los dientes apicectomizados y del tejido alrededor. Los cortes fueron teñidos con la tinción Levai-Laczkó y se realizó el estudio histológico cuantitativo y semicuantitativo, según la norma UNE-EN ISO 10993-6:2007, para medir la presencia de inflamación, fibrosis, infiltrados adiposos y cierre óseo del defecto...

Bacteriologic investigation of the effects of sodium hypochlorite and chlorhexidine during the endodontic treatment of teeth with apical periodontitis

SIQUEIRA JR. et al. Bacteriologic investigation of the effects of sodium hypochlorite and chlorhexidine during the endodontic treatment of teeth with apical periodontitis. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod., v. 104, n. 1, p. 122-130, 2007.

Análise comparativa "in vitro" da extrusão apical de detritos originados por diferentes técnicas de instrumentação do sistema de canais radiculares

Introdução: A extrusão apical detritos (EAD) consequência indesejável da instrumentação canalar pode ser associada a dor/edema, podendo atrasar a cicatrização periapical. O nosso trabalho teve como objectivo avaliar e quantificar a EAD em canais instrumentados por sistemas de instrumentação rotatória contínua e reciprocante. Materiais e Métodos: 80 dentes monocanalares sem tratamento endodôntico prévio foram aleatoriamente divididos em 4 grupos (n=20): One Shape® Protaper® NEXT, Hyflex® EDM e WaveOne® Gold. Um tubo de Eppendorf (TdE) foi pesado antecipadamente numa balança analítica de precisão e com um dente inserido foi montado num dispositivo modificado, similar ao método descrito por Myers & Montgomery. Os canais foram instrumentados e irrigados com água destilada. Os dentes instrumentados foram removidos dos TdE e estes preenchidos com água destilada até perfazer 1,5ml, incubados a 70ºC durante cinco dias sendo pesados novamente, calculando a diferença entre o peso inicial e final determinando o peso dos detritos. Os dados foram analisados estatisticamente utilizando o IBM SPSS Statistics 22, considerando α=0,05. Efetuaram-se testes Kruskal-Wallis e post-hoc com ajustamento do ρ-value pelo método Dunn-Bonferroni. Resultados: Houve EAD em todas as técnicas de instrumentação. A análise estatística mostrou haver diferenças significativas na EAD entre as técnicas utilizadas (α=0,002). Entre as técnicas WaveOne® Gold e One Shape® (α=0,003), WaveOne® Gold e Protaper® NEXT (α=0,023) e WaveOne® Gold e Hyflex® EDM (α=0,028). Conclusões: A técnica One Shape® apresentou menor EAD e a técnica WaveOne® Gold com movimento reciprocante constitui maior fator de risco tendo apresentado maior EAD. Os resultados deste estudo indicam que os profissionais devem estar cientes para a EAD que pode ocorrer com cada instrumento, o que poderá servir de base para a selecção de um instrumento particular. Implicações clínicas: A escolha do sistema de instrumentação canalar influencia a extrusão de detritos. Fontes de financiamento: Agradecimentos as empresas; Micro-Mega, França, COLTÉNE e Dentsply Maillefer, Suíça.

Bacteriologic investigation of the effects of sodium hypochlorite and chlorhexidine during the endodontic treatment of teeth with apical periodontitis

SIQUEIRA JR. et al. Bacteriologic investigation of the effects of sodium hypochlorite and chlorhexidine during the endodontic treatment of teeth with apical periodontitis. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod., v. 104, n. 1, p. 122-130, 2007.