Agglutinating Secretory IgA Preserves Intestinal Epithelial Cell Integrity during Apical Infection by Shigella flexneri.

Shigella flexneri, by invading intestinal epithelial cells (IECs) and inducing inflammatory responses of the colonic mucosa, causes bacillary dysentery. Although M cells overlying Peyer's patches are commonly considered the primary site of entry of S. flexneri, indirect evidence suggests that bacteria can also use IECs as a portal of entry to the lamina propria. Passive delivery of secretory IgA (SIgA), the major immunoglobulin secreted at mucosal surfaces, has been shown to protect rabbits from experimental shigellosis, but no information exists as to its molecular role in maintaining luminal epithelial integrity. We have established that the interaction of virulent S. flexneri with the apical pole of a model intestinal epithelium consisting of polarized Caco-2 cell monolayers resulted in the progressive disruption of the tight junction network and actin depolymerization, eventually resulting in cell death. The lipopolysaccharide (LPS)-specific agglutinating SIgAC5 monoclonal antibody (MAb), but not monomeric IgAC5 or IgGC20 MAbs of the same specificity, achieved protective functions through combined mechanisms, including limitation of the interaction between S. flexneri and epithelial cells, maintenance of the tight junction seal, preservation of the cell morphology, reduction of NF-κB nuclear translocation, and inhibition of proinflammatory mediator secretion. Our results add to the understanding of the function of SIgA-mediated immune exclusion by identifying a mode of action whereby the formation of immune complexes translates into maintenance of the integrity of epithelial cells lining the mucosa. This novel mechanism of protection mediated by SIgA is important to extend the arsenal of effective strategies to fight against S. flexneri mucosal invasion.

Differential regulation of the Hippo pathway by adherens junctions and apical-basal cell polarity modules.

Adherens junctions (AJs) and cell polarity complexes are key players in the establishment and maintenance of apical-basal cell polarity. Loss of AJs or basolateral polarity components promotes tumor formation and metastasis. Recent studies in vertebrate models show that loss of AJs or loss of the basolateral component Scribble (Scrib) cause deregulation of the Hippo tumor suppressor pathway and hyperactivation of its downstream effectors Yes-associated protein (YAP) and Transcriptional coactivator with PDZ-binding motif (TAZ). However, whether AJs and Scrib act through the same or independent mechanisms to regulate Hippo pathway activity is not known. Here, we dissect how disruption of AJs or loss of basolateral components affect the activity of the Drosophila YAP homolog Yorkie (Yki) during imaginal disc development. Surprisingly, disruption of AJs and loss of basolateral proteins produced very different effects on Yki activity. Yki activity was cell-autonomously decreased but non-cell-autonomously elevated in tissues where the AJ components E-cadherin (E-cad) or α-catenin (α-cat) were knocked down. In contrast, scrib knockdown caused a predominantly cell-autonomous activation of Yki. Moreover, disruption of AJs or basolateral proteins had different effects on cell polarity and tissue size. Simultaneous knockdown of α-cat and scrib induced both cell-autonomous and non-cell-autonomous Yki activity. In mammalian cells, knockdown of E-cad or α-cat caused nuclear accumulation and activation of YAP without overt effects on Scrib localization and vice versa. Therefore, our results indicate the existence of multiple, genetically separable inputs from AJs and cell polarity complexes into Yki/YAP regulation.

A fully echo-guided trans-apical aortic valve implantation.

The trans-apical aortic valve implantation (TA-AVI) is an established technique for high-risk patients requiring aortic valve replacement. Traditionally, preoperative (computed tomography (CT) scan, coronary angiogram) and intra-operative imaging (fluoroscopy) for stent-valve positioning and implantation require contrast medium injections. To preserve the renal function in elderly patients suffering from chronic renal insufficiency, a fully echo-guided trans-catheter valve implantation seems to be a reasonable alternative. We report the first successful TA-AVI procedure performed solely under trans-oesophageal echocardiogram control, in the absence of contrast medium injections.

Simptomatologia i microorganismes associats a la necrosi bruna apical de la noguera (Juglans regia)

La Necrosi Apical Bruna (BAN, brown apical necrosis, segons les sigles en anglès)Es va detectar per primer cop l’any 1997 a Extremadura degut a la severa caiguda defruits. Avui dia la malaltia és present a quasi totes les zones productores de lamediterrània. Els símptomes difereixen dels provocats per Xanthomonas arboricola pv.juglandis i Gnomonia leptsostyla.. S’observa que els fruits afectats presenten una tacabruna a la zona apical i necrosi dels teixits interiors. El grup de Patologia Vegetal de laUniversitat de Girona participa i dirigeix la tasca sobre l’etiologia de la BAN dins laxarxa europea d’investigació en bacteris patògens de fruiters ,COST873. Hi ha una certacontrovèrsia en la definició dels símptomes i agents causals. Tots els grups coincideixenen afirmar que es tracta d’una malaltia complexa amb diferents organismes implicats

Sorting mechanisms regulating membrane protein traffic in the apical transcytotic pathway of polarized MDCK cells.

The transcytotic pathway followed by the polymeric IgA receptor (pIgR) carrying its bound ligand (dIgA) from the basolateral to the apical surface of polarized MDCK cells has been mapped using morphological tracers. At 20 degreesC dIgA-pIgR internalize to interconnected groups of vacuoles and tubules that comprise the endosomal compartment and in which they codistribute with internalized transferrin receptors (TR) and epidermal growth factor receptors (EGFR). Upon transfer to 37 degreesC the endosome vacuoles develop long tubules that give rise to a distinctive population of 100-nm-diam cup-shaped vesicles containing pIgR. At the same time, the endosome gives rise to multivesicular endosomes (MVB) enriched in EGFR and to 60-nm-diam basolateral vesicles. The cup-shaped vesicles carry the dIgA/pIgR complexes to the apical surface where they exocytose. Using video microscopy and correlative electron microscopy to study cells grown thin and flat we show that endosome vacuoles tubulate in response to dIgA/pIgR but that the tubules contain TR as well as pIgR. However, we show that TR are removed from these dIgA-induced tubules via clathrin-coated buds and, as a result, the cup-shaped vesicles to which the tubules give rise become enriched in dIgA/pIgR. Taken together with the published information available on pIgR trafficking signals, our observations suggest that the steady-state concentrations of TR and unoccupied pIgR on the basolateral surface of polarized MDCK cells are maintained by a signal-dependent, clathrin-based sorting mechanism that operates along the length of the transcytotic pathway. We propose that the differential sorting of occupied receptors within the MDCK endosome is achieved by this clathrin-based mechanism continuously retrieving receptors like TR from the pathways that deliver pIgR to the apical surface and EGFR to the lysosome.

Transapical aortic valve replacement through a chronic apical aneurysm.

Transapical aortic valve replacement through an apical aneurysm is traditionally contraindicated because of the risk of severe systemic embolization when thrombi are present. However, a chronic fibrotic aneurysm without apical thrombi carries a low risk of distal embolization and can be safely employed for a transapical transcatheter aortic valve replacement in case of absence of an alternative access site (severe vascular disease, small vascular sizes and diseased calcified aorta). We illustrate our experience with a 73-year-old patient suffering from symptomatic aortic valve stenosis, coronary artery disease with occluded left anterior descending artery, left ventricular apical aneurysm and severe peripheral vascular disease, who successfully underwent a transapical 26 mm Sapien? XT stent-valve implantation through the fibrotic thin akinetic apical wall.

Abnormal apical-to-basal transport of dietary ovalbumin by secretory IgA stimulates a mucosal Th1 response.

In celiac disease, enhanced permeability to gliadin peptides can result from their apico-basal transport by secretory immunoglobulin A1 (SIgA1) binding to the CD71 receptor ectopically expressed at the gut epithelial surface. Herein, we have established a mouse model in which there is apico-basal transport of the model antigen ovalbumin (OVA) by specific SIgA1 and have analyzed local T-cell activation. Transgenic DO11.10 mice were grafted with a hybridoma-secreting OVA-specific humanized IgA1, which could bind mouse CD71 and which were released in the intestinal lumen as SIgA. CD71 expression was induced at the gut apical surface by treating the mice with tyrphostin A8. Following gavage of the mice with OVA, OVA-specific CD4(+) T cells isolated from the mesenteric lymph nodes displayed higher expression of the activation marker CD69 and produced more interferon gamma in mice bearing the hybridoma-secreting OVA-specific IgA1, than in ungrafted mice or in mice grafted with an irrelevant hybridoma. These results indicate that the protective role of SIgA1 might be jeopardized in human pathological conditions associated with ectopic expression of CD71 at the gut surface.

A new training set-up for trans-apical aortic valve replacement.

Trans-apical aortic valve replacement (AVR) is a new and rapidly growing therapy. However, there are only few training opportunities. The objective of our work is to build an appropriate artificial model of the heart that can replace the use of animals for surgical training in trans-apical AVR procedures. To reduce the necessity for fluoroscopy, we pursued the goal of building a translucent model of the heart that has nature-like dimensions. A simplified 3D model of a human heart with its aortic root was created in silico using the SolidWorks Computer-Aided Design (CAD) program. This heart model was printed using a rapid prototyping system developed by the Fab@Home project and dip-coated two times with dispersion silicone. The translucency of the heart model allows the perception of the deployment area of the valved-stent without using heavy imaging support. The final model was then placed in a human manikin for surgical training on trans-apical AVR procedure. Trans-apical AVR with all the necessary steps (puncture, wiring, catheterization, ballooning etc.) can be realized repeatedly in this setting.

The apical localization of transcribing RNA polymerases on supercoiled DNA prevents their rotation around the template.

The interaction of Escherichia coli RNA polymerase with supercoiled DNA was visualized by cryo-electron microscopy of vitrified samples and by classical electron microscopy methods. We observed that when E. coli RNA polymerase binds to a promoter on supercoiled DNA, this promoter becomes located at an apical loop of the interwound DNA molecule. During transcription RNA polymerase shifts the apical loop along the DNA, always remaining at the top of the moving loop. This relationship between RNA polymerase and the supercoiled template precludes circling of the RNA polymerase around the DNA and prevents the growing RNA transcript from becoming entangled with the template DNA.

Feasibility of transapical aortic valve replacement through a left ventricular apical diverticulum.

ABSTRACT: Transapical aortic valve replacement is an established technique performed in high-risk patients with symptomatic aortic valve stenosis and vascular disease contraindicating trans-vascular and trans-aortic procedures. The presence of a left ventricular apical diverticulum is a rare event and the treatment depends on dimensions and estimated risk of embolisation, rupture, or onset of ventricular arrhythmias. The diagnosis is based on standard cardiac imaging and symptoms are very rare. In this case report we illustrate our experience with a 81 years old female patient suffering from symptomatic aortic valve stenosis, respiratory disease, chronic renal failure and severe peripheral vascular disease (logistic euroscore: 42%), who successfully underwent a transapical 23 mm balloon-expandable stent-valve implantation through an apical diverticulum of the left ventricle. Intra-luminal thrombi were absent and during the same procedure were able to treat the valve disease and to successfully exclude the apical diverticulum without complications and through a mini thoracotomy. To the best of our knowledge, this is the first time that a transapical procedure is successfully performed through an apical diverticulum.

Infiltração de água em um Latossolo vermelho-escuro sob condições de chuva intensa em diferentes sistemas de manejo

A infiltração de água é uma propriedade que reflete as condições físicas do solo, principalmente quanto à sua qualidade estrutural. Foi realizado um estudo na Embrapa Trigo, em Passo Fundo (RS), em um Latossolo Vermelho-Escuro, submetido por longo tempo a diferentes sistemas de manejo, com o objetivo de verificar a influência desses sistemas na taxa de infiltração de água, considerando as diferentes coberturas de solo e condições físicas gerais por eles propiciadas. A área experimental estava cultivada com milho, tendo-se aplicado chuva simulada de 120 mm h-1, durante 90 minutos, em três repetições, em três épocas distintas: (a) 45 dias após a semeadura do milho; (b) logo após a colheita do milho e (c) logo após a semeadura de aveia. As chuvas simuladas foram aplicadas sobre parcelas de 0,81 m² distribuídas dentro de macroparcelas com os sistemas de manejo em preparo convencional, cultivo mínimo e sistema plantio direto. O cultivo mínimo apresentou, em todas as épocas, as maiores taxas de infiltração de água no solo, e o sistema plantio direto produziu as maiores quantidades de palha na superfície do solo.

Infiltração de água em um podzólico vermelho-escuro sob dois métodos de preparo, usando chuva simulada com duas intensidades

Realizou-se no campo, num Podzólico Vermelho-Escuro, um experimento, para estudar a taxa de infiltração, tempos de empoçamento e início de escoamento superficial de água, sob preparo convencional e plantio direto, usando chuva simulada de intensidades médias constantes de 63,0 e 87,0 mm h-1. Foi usado um microssimulador de chuvas com bicos Veejet 80-100 e 80-150, sendo a área de observação para os testes de infiltração de 0,82 m². O modelo de Smith foi empregado para descrever o processo de infiltração. Foram usadas parcelas subdivididas com quatro repetições, cultivadas com aveia (Avena strigosa) e soja (Glycine max). Os tempos de empoçamento e de início de escoamento diminuíram com o aumento da intensidade de chuva. Considerando os tipos de preparos, os tempos foram menores no plantio direto. A infiltração acumulada e a taxa constante de infiltração foram maiores no plantio direto. Neste preparo, não houve diferença na taxa constante para as duas intensidades; para o preparo convencional, a taxa foi menor sob a chuva de maior intensidade. O parâmetro b do modelo teve pequena variação e a taxa constante de infiltração foi superestimada.

Infiltração de água e perdas de água e solo por erosão influenciadas por diferentes métodos de melhoramento da pastagem nativa gaúcha

A paralisação do crescimento da pastagem nativa no período do inverno na região Sul do Brasil tem incentivado técnicas de melhoramento das pastagens. Com o objetivo de estudar a infiltração de água no solo e as perdas de solo e água por erosão influenciadas por métodos de melhoramento da pastagem nativa, realizou-se um estudo em área da Estação Experimental Agronômica da UFRGS, no município de Eldorado do Sul (RS), em um Podzólico Vermelho-Amarelo submetido ao uso prolongado com pastagem nativa, no qual se fez a introdução de uma mistura das espécies hibernais: aveia preta (Avena strigosa ), azevém (Lolium multiflorum ) e trevo vesiculoso (Trifolium vesiculosum). O delineamento experimental foi completamente casualizado e com cinco tratamentos que diferiram quanto à maneira de introdução das novas espécies: testemunha (a lanço), gradagem, plantio direto, convencional e subsolagem. As parcelas tinham a dimensão de 3,5 x 11,0 m e uma declividade aproximada de 0,107 m m-1. Aplicaram-se chuvas simuladas de 64 mm h-1, durante 75 min, em três épocas: (a) aos 55 dias do preparo do solo e semeadura, logo após o primeiro pastejo, (b) aos 125 dias do preparo do solo e semeadura, e (c) logo após o segundo pastejo, aos 175 dias do preparo do solo e semeadura. Entre as chuvas simuladas, realizou-se um pastejo por dois dias. A subsolagem apresentou a maior taxa constante de infiltração e a menor taxa constante de enxurrada. As perdas de solo foram pequenas nas três épocas avaliadas. As maiores perdas de água ocorreram na testemunha e as menores na subsolagem, indicando ter sido este último tratamento efetivo em quebrar camadas compactadas subsuperficiais.

Influência da crosta superficial nas taxas de infiltração de água medidas com simulador de chuva e anéis concêntricos

A taxa de infiltração de água no solo, conjuntamente com a taxa de chuva, determina a taxa de enxurrada, constituindo medida importante para a previsão de erosão e estudos de conservação do solo. Pode-se medir a taxa de infiltração por meio de um infiltrômetro de cilindros concêntricos (CC) ou de um simulador de chuva (SC). A utilização do CC requer menos trabalho do que o SC, mas os resultados obtidos com o CC comumente estão em desacordo com as taxas de infiltração que ocorrem em situações de chuva natural, possivelmente por causa da formação de uma crosta superficial sob chuva. A formação dessa crosta procede tanto de fatores independentes do manejo do solo, como a intensidade da chuva e a textura do solo, quanto de fatores dependentes do manejo, principalmente a resistência ao cisalhamento e a cobertura vegetal. Em situações favoráveis à ocorrência de formação de crosta, espera-se que taxas de infiltração medidas com o CC superestimem as taxas reais durante uma chuva. Para verificar a correlação entre o manejo do solo e a razão das taxas de infiltração obtidas com CC e SC, essas taxas foram medidas por ambos os métodos em dois Podzólicos (Ultissolos) da região sul do Brasil. Os tratamentos diferiram com a cobertura vegetal e com a existência de crosta superficial. Medidas da taxa de infiltração com SC foram realizadas com um simulador de chuva com bicos aspersores Veejet 80100, em parcelas de 3,5 x 11,0 m, gerando chuva de intensidade de 64,2 mm h-1, durante 100 a 150 minutos. As parcelas foram delimitadas com chapas de metal e a enxurrada foi coletada no lado inferior por três segundos a cada três minutos. O CC consistiu de dois cilindros (diâmetros de 0,30 e 0,60 m), cravados no solo até 0,10 m pouco antes da determinação com o SC. A área dentro dos anéis foi inundada e as taxas de infiltração foram registradas dentro do anel interior, em intervalos de tempo regulares, com três repetições por parcela. Os resultados mostraram que a taxa de infiltração final, quando medida com o CC, foi de cinco a 10 vezes superior à medida com o SC. Medidas da taxa de infiltração com o CC diferiram menos das do SC, quando uma cobertura do solo estava presente. Taxas instantâneas de infiltração diferiram menos quando a crosta preexistente foi removida anterior aos testes. Esses resultados mostram que as taxas de infiltração determinadas pelo CC e SC não podem ser utilizadas em estudos de conservação do solo sem correção e que essa correção deve considerar as condições de manejo, especialmente aquelas relacionadas com a existência e formação de crosta superficial e com a presença de cobertura vegetal.