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...

Stimulation of GABAergic neurons of the lateral septum and its effect on movement speed

The lateral septum is associated with the regulation of innate behavior, motivation, and locomotion. Its complex interconnections with cognitive and affective regions such as the hippocampus, hypothalamus, and medial septum have made it an attractive region for studying how motivation regulates behavior in context-specific settings. This GABAergic brain region’s main output is the lateral hypothalamus, which provides downstream signaling of motor commands. Even though stimulation of lateral septum projections to the hypothalamus have shown to decrease running speed in free behaving mice, characterizing movement kinematics due to LS activation has not been studied. GABAergic medium spiny neurons of the lateral septum were selectively activated through the use of optogenetic techniques in transgenic mice. Photostimulation of the lateral septum at theta frequencies caused a non-significant decrease in head and back speed. 3D motion analysis of body movement under photostimulation was quantified, revealing a slow, linear decrease of body speed as photostimulation progressed. These results support the role of lateral septum activation in movement regulation and shed light on the specific manner in which stimulation of the LS gradually decreases movement speed.

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.

Determinación del tamaño de la preparación apical: un dilema clínico en endodoncia

La determinación del tamaño de la preparación apical es un tema de constante debate, ya que a pesar de la evidencia científica disponible aún no se logra un consenso para saber cuál es el diámetro adecuado para obtener un tratamiento exitoso; en este sentido, el clínico debe ser crítico ante cada tratamiento, evaluando el diagnóstico y la anatomía del sistema de conductos radiculares. En la presente revisión, se realizó una búsqueda de la literatura disponible, con el fin de ofrecer al odontólogo un enfoque ante sus procedimientos clínicos, considerando diferentes parámetros, así como las distintas implicaciones clínicas que derivan de la determinación del tamaño de la preparación apical, lo cual constituye un aspecto importante dentro del proceso de la preparación biomecánica.

Reabsorción radicular externa apical en incisivos y caninos superiores e inferiores sometidos a tratamiento de Ortodoncia en etapa inicial

La Reabsorción radicular externa apical es un proceso inflamatorio común asociado con el tratamiento de ortodoncia. El objetivo de este estudio fue comparar la magnitud de la RREA de los incisivos y caninos superiores e inferiores en pacientes sometidos a la fase inicial del tratamiento de ortodoncia con tres diferentes tipos de brackets. La muestra consistió en 23 casos de pacientes con edades comprendidas entre 12 y 27 años, fueron divididos aleatoriamente en 3 grupos. Grupo I (n=9; 5 mujeres y 4 varones), utilizando brackets pasivos de autoligado Damon Q Grupo II (n= 8; 4 mujeres y 4 varones) utilizando brackets convencionales prescripción Roth y MBT. Grupo III (n=6; 4 mujeres y 2 varones) utilizando brackets Biofuncional QR. Se analizaron 264 dientes (caninos e incisivos superiores e inferiores) mediante el uso de tomografía computarizada de haz cónico (NewTom VGi Cone Beam 3D Imaging), con el escáner NNT Viewer (versión 4.6 NewTom). Los resultados obtenidos indican que la media de reabsorción se mostró distinta para los diferentes protocolos de tratamiento, así como diferente de acuerdo a la pieza analizada, sin embargo, la prueba de ANOVA no encontró diferencias estadísticamente significativas al comparar la media de reabsorción de cada pieza por tipo de brackets empleados o tratamiento realizado (p >0,05). Se concluye que la RREA se ha producido en todos los dientes evaluados, durante los 6 primeros meses de tratamiento y el diseño del bracket (autoligado pasivo, convencional o QR) no influye en el grado de reabsorción radicular.

Persistent Truncus Arteriosus With Intact Ventricular Septum: Clinical, Hemodynamic and Short-term Surgical Outcome

Introduction: Truncus arteriosus with intact ventricular septum is a rare and unique variant of persistent truncus arteriosus (PTA) which usually presents with central cyanosis and congestive heart failure in neonate and early infancy. Associated cardiac and non-cardiac anomalies may affect morbidity and mortality of these patients. Case Presentation: We describe clinical presentation, echocardiography and angiographic features of a 7-month old boy with PTA and intact ventricular septum who underwent surgical repair of the anomaly at our institution. Operative findings, surgical procedure and short-term outcome are reported. Conclusions: While our patient had systemic pulmonary arterial pressure at the time of complete surgical repair, it was improved after surgery.